Heterocyclic compounds as btk inhibitors

ABSTRACT

The present invention relates to heterocyclic compounds or their pharmaceutically acceptable salts thereof as inhibitors of Bruton&#39;s tyrosine kinase (BTK) and its C481 mutant. The present invention relates to compounds shown in Formula (I) and their pharmaceutically acceptable salts thereof. The present invention also relates to methods for preparing compounds or their pharmaceutically acceptable salts thereof. Compounds of the present invention can be used to treat and/or prevent related diseases mediated by BTK or its C481 mutant, especially cancer and autoimmune diseases.

This application is a continuation of PCT/CN2021/113535, filed Aug. 19,2021, which claims the priority of Chinese Application No.202010836243.4, filed Aug. 20, 2020. The contents of theabove-identified applications are incorporated herein by reference intheir entireties.

TECHNICAL FIELD

The invention relates to heterocyclic compounds or theirpharmaceutically acceptable salts thereof, suitable for regulating orinhibiting activities of Bruton tyrosine kinase (BTK) and its C481mutant. The present invention also relates to methods for preparing thecompounds or their pharmaceutically acceptable salts thereof. Thepresent invention further relates to the uses and methods of use of thecompounds or their pharmaceutically acceptable salts thereof in thetreatment and/or prevention of cancer and autoimmune diseases.

BACKGROUND ART

BTK is an important non-receptor tyrosine kinase that mediates cellsignal transduction, which exists in plasma cells including B-cells.B-cells are activated through B-cell receptor (BCR) and BTK plays animportant role in the BCR-mediated signaling pathway. After BCR onB-cells is activated, it causes the activation of BTK which leads to anincrease in the concentration of downstream phospholipase C (PLC) andactivates the IP3 and DAG signaling pathways. This signaling pathway canpromote cell proliferation, adhesion and survival, and plays animportant role in the development of B-cell lymphoma.

BTK inhibitors inhibit the proliferation of B lymphoma cells byinhibiting the activity of BTK, destroy adhesion of tumor cells, andpromote tumor cell apoptosis, making BTK a compelling drug target forB-cell related cancers, such as non-Hodgkin's lymphoma (NHL), chroniclymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL), mantle celllymphoma (MCL), Waldenstrom's macroglobulinemia (WM), marginal zoneLymphoma (MZL), central nervous system leukemia (CNSL), etc. Several BTKinhibitors are currently on the market, including Abbvie/JNJ'sibrutinib, AZ's acalabrutinib, Beigene's zanubrutinib and Gilead/Ono'stirabrutinib, and more BTK inhibitors are in clinical research.

In addition to treating B-cell associated lymphomas, BTK inhibitors canalso inhibit the production of B-cell autoantibodies and cytokines. Inautoimmune diseases, B-cells present their own antigens, promote T-cellactivation, secrete inflammatory factors that cause tissue damage, andat the same time activate B-cells to produce a large number ofantibodies to trigger an autoimmune response. T- and B-cells interact toeach other to form a positive feedback regulatory chain which leads touncontrolled autoimmune responses and aggravates tissue pathologicaldamage. Studies have shown that there are regulatory B-cells in the bodywhich can negatively regulate the immune response and inhibitimmune-mediated inflammation through the secretion of interleukin 10(IL-10) or transforming growth factor β1 (TGF-β1) and other mechanisms.Therefore, BTK can be a drug target for autoimmune diseases, such asrheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupuserythematosus (SLE), pemphigus, etc. For autoimmune indications, BTKinhibitors are still in clinical research. Among them, Sanofi'srilzabrutinib and Merck Serono's evobrutinib have achieved effectiveresults in the treatment of pemphigus and multiple sclerosis,respectively.

Most of BTK inhibitors on the market and under research are irreversibleinhibitors which inhibit the activity of BTK by covalently binding tothe cysteine residue located at 481 of the BTK protein. After someB-cell lymphoma patients received ibrutinib treatment for a period oftime, BTK's C481 mutation, such as C481S, made ibrutinib lose itscovalent binding point with the protein, resulting in a decrease in theactivity of ibrutinib, thereby making patients resistant to theibrutinib treatment (Quinquenel, et. al Blood 2019, 134, 641-644). Thereexists a need for BTK inhibitors which effectively inhibit theactivities of BTK and its C481 mutant, thereby overcoming the drugresistance caused by the C481 mutation associated with irreversible BTKinhibitors.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless otherwise stated, the following terms used in this applicationhave the following meanings.

“CSF/plasma ratio (Kp)” refers to the ratio of a compound concentrationin cerebrospinal fluid (CSF) vs. in plasma. The ability of a compound tocross blood-brain barrier (BBB) is assessed by measuring itsconcentrations in CSF and plasma in rodents, and determining the ratio(Kp, CSF).

“C_(x-y)” refers to a range of the number of carbon atoms, where x and yare both integers, for example, C₃₋₈ cycloalkyl stands for cycloalkylhaving 3 to 8 carbon atoms.

“Alkyl” refers to a saturated straight-chain or branched-chainhydrocarbyl substituent containing 1 to 20 carbon atoms, for example, 1to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms.Unrestricted examples of alkyl include but are not limited to methyl,ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl,n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl,1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl,1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl,1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl and2-ethylbutyl.

“Cycloalkyl” refers to a saturated cyclic hydrocarbyl substituentcontaining 3 to 14 annular carbon atoms. Cycloalkyl may be a mono carbonring substituent, typically containing 3 to 8, 3 to 7, or 3 to 6 carbonatoms. Unrestricted examples of monocyclic cycloalkyl include but arenot limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl andcycloheptyl. Cycloalkyl may also be a substituent with two or three monorings that are fused together, such as decahydronaphthyl,bicyclo[2.2.2]octane and spiro[3.3]heptane.

“Heterocyclyl or heterocycle” refers to a saturated or partiallyunsaturated monocyclic or polycyclic group containing 3 to 20 annularatoms, for example, 3 to 14, 3 to 12, 3 to 10, 3 to 8, 3 to 6, or 5 to 6annular atoms in which one or more of the annular atoms are selectedfrom N, O and S(O)_(m) (where m is an integer from 0 to 2). Preferably,it may have 3 to 12 annular atoms, more preferably 3 to 10 ring atoms,more preferably 4 to 7 ring atoms, more preferably 4 to 6 ring atoms,most preferably 5 or 6 ring atoms, wherein 1 to 4 are heteroatoms, 1 to3 are heteroatoms, or 1 to 2 are heteroatoms. Unrestricted examples ofmonocyclic heterocyclyl include but are not limited to pyrrolidinyl,oxetanyl, piperidyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl,tetrahydrothiopyranyl, morpholinyl, thiomorpholinyl, homopiperazinyl andazetidinyl. Polycyclic heterocyclyl includes fused, bridged or spiropolycyclic heterocycle, such as octahydrocyclopenta[c]pyrrole,octahydropyrrole[1,2-a]pyrazine, 3,8-diazabicyclo[3.2.1]octane,5-azaspiro[2.4]heptane and 2-oxa-7-azaspiro[3.5]nonane.

“Aryl or aryl ring” refers to an aromatic monocyclic or fused polycyclicgroup containing 6 to 14 carbon atoms, preferably 6- to 10-membered,such as phenyl and naphthyl, most preferably phenyl. The aryl ring maybe fused with a heteroaryl, heterocyclic or cycloalkyl ring.Unrestricted examples include but are not limited to:

“Heteroaryl or heteroaryl ring” refers to a heteroaromatic systemcontaining 5 to 14 annular atoms, of which 1 to 4 annular atoms areselected from heteroatoms including O, S and N. Heteroaryl preferably is5- to 10-membered, and more preferably 5- or 6-membered, such as furyl,thienyl, pyridyl, pyrrolyl, pyrimidyl, pyrazinyl, pyrazolyl, imidazolyl,tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolinyl,isoquinolinyl, indolyl and isoindolyl. The heteroaryl ring may be fusedwith an aryl, heterocyclyl or cycloalkyl ring. Unrestricted examplesinclude but are not limited to:

“Halogen” refers to F, Cl, Br, or I.

“Cyano” refers to —CN.

“Optional” means that the event or environment described later may butneed not occur, and the expression includes the occurrence ornon-occurrence of the event or environment. For example, “heterocyclicoptionally substituted by an alkyl group” includes the case where theheterocyclic is substituted by an alkyl group and the case where theheterocyclic is not substituted by an alkyl group.

“Substitution” refers to one or more hydrogen atoms in a group, forexample, 1 to 3 hydrogen atoms that are independently substituted by acorresponding number of substituents. The substituents are located onlyin their possible chemical positions understood by those skilled in theart. The substituents include but are not limited to halogen, hydroxyl,cyano, nitro, oxo, —SFS, C₁₋₄ alkyl, C₁₋₄ alkoxyl, etc.

“Isomers” refer to compounds that have the same molecular formula, buttheir atomic binding position or spatial arrangement is different.Isomers with different arrangement of their atoms in space are called“stereoisomers”. Stereoisomers include optical isomers, geometricisomers, and conformational isomers.

Compounds of the present invention can exist as optical isomers. Opticalisomers include enantiomers and diastereomers. An enantiomer is one oftwo stereoisomers that are mirror images of each other and arenon-superposable. A racemic mixture or racemate is one that has equalamounts of left- and right-handed enantiomers of a chiral molecule.Diastereomers are stereoisomers that are not mirror images of oneanother and are non-superimposable on one another. When a compound is asingle isomer and its absolute configuration is determined, it isreferred as a “R” or “S” isomer according to the configuration of thesubstituents around the chiral carbon atom. When its absoluteconfiguration is not determined, it is referred as a (+) or (−) isomeraccording to its measured optical rotation value. Methods for preparingand separating optical isomers are known to those skilled in the art.

Compounds of the present invention may also have geometric isomersresulting from the distribution of substituents around carbon-carbondouble bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclylgroups. The substituents around the carbon-carbon double bond orcarbon-nitrogen bond are designated to be in a Z or E configuration, andthe substituents around the cycloalkyl or heterocycle are designated tobe in a cis or trans configuration.

Compounds of the present invention may also show tautomerism, such asketo-enol tautomerism.

The present invention includes any tautomeric or stereoisomeric formsand mixtures thereof and is not limited to any tautomeric orstereoisomeric forms used in the compound nomenclature or chemicalstructural formulae.

“Isotopes” include all stable isotopes of the atoms appearing in thecompounds of the present invention. Isotopes include those atoms withthe same atomic number but in different masses. Examples of isotopessuitable for incorporation into the compounds of the present inventionare isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorineand chlorine, for example but not limited to ²H (D), ³H, ¹³C, ¹⁴C, ¹⁵N,¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F and ³⁶Cl. The isotopically labeledcompounds of the present invention can generally be prepared byconventional techniques known to those skilled in the art or by methodssimilar to those described in the embodiments using appropriateisotopically labeled reagents instead of non-isotopically labeledreagents. Such compounds have various potential uses, for example, asstandards and reagents in the determination of biological activities. Inthe case of stable isotopes, such compounds have the potential tobeneficially alter biological, pharmacological, or pharmacokineticproperties. Deuterium ²H (D) is a preferable isotope of the presentinvention. For example, hydrogen in methyl, methylene or methine can bereplaced by deuterium.

Compounds of the present invention can be administered in form ofprodrugs. “Prodrugs” refer to derivatives that are converted intobiologically active compounds under the physiological condition in vivo,for example, by oxidation, reduction, and hydrolysis (each of whichoccurs with or without the participation of enzymes). Examples of aprodrug are a compound of the present invention in which an amino isacylated, alkylated or phoshorylated, for example eicosanoyl amino,alanyl amino and pivaloyloxymethyl amino; a hydroxyl is acylated,alkylated or phoshorylated or converted into borate, for exampleacetoxy, palmitoyloxy, pivaloyloxy, succinyloxy, fumaroyloxy andalanyloxy; a carbonyl is esterified or amidated; and a thiol forms adisulfide bridge with a carrier molecule that selectively delivers thedrug to the target and/or to the cytosol of cells, such as peptide.Prodrugs may be prepared from the compounds of the present inventionaccording to well-known methods.

“Pharmaceutically acceptable salts” refer to the salts made fromcompounds of the present invention with pharmaceutically acceptablebases or acids, including inorganic alkalis or acids and organic basesor acids, under the condition that the compounds contain one or moreacidic or basic groups. Compounds of the present invention that containacidic groups can exist in form of salts, for example, as alkali metalsalts, alkaline earth metal salts, or ammonium salts. For example, suchsalts include sodium salts, potassium salts, calcium salts, magnesiumsalts or ammonia or organic amine salts such as salts of ethylamine,ethanolamine, triethanolamine or amino acids. Compounds of the presentinvention that contain basic groups can exist in form of salts asinorganic or organic acid salts. Examples of suitable acids includehydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid,nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid,salicylic acid, benzoic acid, formic acid, propanoic acid, pivalic acid,malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid,malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbicacid, isonicotinic acid, citric acid, adipic acid and other acids knownto those skilled in the art. If compounds of the present inventioncontain both acidic and basic groups in the molecule, the presentinvention further includes internal salts in addition to the mentionedsalt forms. Each salt can be obtained by conventional methods known tothose skilled in the art, for example by mixing a compound of thepresent invention with an organic or inorganic acid or base in a solventor dispersant, or by anion exchange or cation exchange with anothersalt.

“Pharmaceutical composition” refers to a composition containing one ormore of compounds of the present invention or their pharmaceuticallyacceptable salts, stable isotope derivatives, isomers, prodrugs, andmixtures thereof, and other components such as pharmaceuticallyacceptable carrier and excipients.

“Cancer or lymphoma or leukemia” includes but is not limited to B-cellmalignancies, B-cell lymphoma, diffuse large B-cell lymphoma, chroniclymphocytic leukemia, small lymphocytic lymphoma, non-Hodgkin Lymphoma(such as ABC-DLBCL), mantle cell lymphoma, follicular lymphoma,Waldenstrom's macroglobulinemia, marginal zone lymphoma, central nervoussystem lymphoma, chronic lymphocytic lymphoma, B-cell prelymphocyticleukemia, plasma cell lymphoma, multiple myeloma, various solid tumors(such as melanoma, bone cancer, brain cancer, colon cancer, livercancer, skin cancer, kidney cancer, lung cancer, muscle cancer, bladdercancer, digestive tract/stomach Intestinal cancer, breast cancer,ovarian cancer, head and neck cancer, prostate cancer), etc.

“Autoimmune or inflammatory disease” includes but is not limited toarthritis, multiple sclerosis, osteoporosis, inflammatory bowel disease,colitis, Crohn's disease, lupus, rheumatoid Arthritis, psoriaticarthritis, lupus nephritis, Sjogren's syndrome, IgG4-related diseases,idiopathic thrombocytopenic purpura, immune thrombocytopenia, Wright'ssyndrome, psoriasis, Behcet's disease, asthma, Pemphigus, diabetes,myasthenia gravis, Guillain-Barre syndrome, Graves' disease, Hashimoto'sthyroiditis, vasculitis, autoimmune vasculitis, granuloma with multiplevasculitis, autoimmune hepatitis, etc.

“Therapeutically effective amount” refers to an amount of compounds ofthe present invention that can effectively inhibit activities of BTK andits C481 mutant, and/or treat or prevent the diseases mediated by BTKand its C481 mutant.

“Patients” refer to mammals, preferably humans.

The present invention are directed to compounds with a structure asshown in Formula (I) as reversible BTK inhibitors, which effectivelyinhibit the activities of BTK and its C481 mutant, thereby overcomingthe drug resistance caused by the C481 mutation associated withirreversible BTK inhibitors.

The present invention provides heterocyclic compounds as shown inFormula (I) or their pharmaceutically acceptable salts, stable isotopederivatives, isomers, and prodrugs thereof,

where:

A is

where → indicates that A is connected to the benzene ring and

indicates that A is connected to E;

Ring K is phenyl or pyridyl, where the phenyl and pyridyl are optionallysubstituted by one or more substituents selected from halogen, cyano,C₁₋₆ alkyl or —OR^(a);

E is C₁₋₆ alkyl, C₃₋₁₀ cycloalkyl, 3-10 membered heterocyclyl, C₆₋₁₀aryl or 5-10 membered heteroaryl, where the alkyl, cycloalkyl,heterocyclyl, aryl and heteroaryl are optionally substituted by one ormore substituents selected from D, halogen, cyano, —OR^(b),—NR^(b)R^(c), —COOR^(b), —C(O)R^(b), —C(O)NR^(b)R^(c) or R^(e);

R^(e) is C₁₋₆ alkyl, C₃₋₁₀ cycloalkyl or 3-10 membered heterocyclyl,where one or more hydrogens of the alkyl, cycloalkyl and heterocyclylare optionally substituted by halogen, cyano, —OR^(b), —NR^(b)R^(c),—COOR^(b), —C(O)R^(b) or —C(O)NR^(b)R^(c);

R¹ is H, halogen, —OR^(a) or C₁₋₆ alkyl;

R^(a) is C₁₋₆ alkyl, where one or more hydrogens of the alkyl areoptionally substituted by D or fluorine; and

R^(b) and R^(c) are each independently selected from H, C₁₋₆ alkyl, C₃₋₆cycloalkyl or 4-6 membered heterocyclyl.

In one embodiment, A is

In one embodiment, A is

In another embodiment, A is

In another embodiment, A is

In another embodiment, A is

In one embodiment, E is C₁₋₆ alkyl, wherein one or more hydrogens of thealkyl are optionally substituted by D or fluorine, for example—CH(CH₃)(CF₃).

In one embodiment, E is C₃₋₇ monocyclic cycloalkyl or 4-8 memberedmonocyclic heterocyclyl containing N and/or O. For example, E is

In one embodiment, E is C₅₋₁₀ polycyclic cycloalkyl or 5-10 memberedpolycyclic heterocyclyl containing O. For example, E is

The monocyclic cycloalkyl, polycyclic cycloalkyl, monocyclicheterocyclyl, and polycyclic heterocyclyl are independently andoptionally substituted by one or more substituents selected from D,halogen, —OR^(b), —NR^(b)R^(c), —COOR^(b), —C(O)R^(b), —C(O)NR^(b)R^(c)or C₁₋₆ alkyl, where one or more hydrogens of the alkyl are furtheroptionally substituted by halogen, —OR^(b) or —NR^(b)R^(c); R^(b) andR^(c) are each independently selected from H, C₁₋₆ alkyl, C₃₋₆cycloalkyl or 4-6 containing N and/or O containing heterocyclic group(e.g., morpholinyl).

In a preferred embodiment, E is

G is H, fluorine, —OR^(b), —NR^(b)R^(c), —COOR^(b), —C(O)R^(b),—C(O)NR^(b)R^(c) or C₁₋₂ alkyl, where one or more hydrogens of the alkylgroup are optionally substituted by fluorine, —OH or —NH₂; R^(b) andR^(c) are each independently selected from H, C₁₋₂ alkyl, C₃₋₆cycloalkyl or N and/or O containing 4-6 membered heterocyclyl (forexample, morpholinyl).

In a preferred embodiment, ring K is phenyl, where the phenyl isoptionally substituted by one or two substituents selected from halogenor —OR^(a); R^(a) is C₁₋₂ alkyl, where one or more hydrogens of thealkyl is optionally substituted by D.

For example, ring K is

R^(a) is a C₁₋₂ alkyl group, where one or more hydrogens of the alkylare optionally substituted by D.

In one embodiment, R¹ is H.

In another embodiment, R¹ is halogen.

In one embodiment, the present invention relates to compounds that arehave a substantial brain penetration and have a blood-brain barrierpermeability.

In one embodiment, the present invention relates to CNS penetrantcompounds that have CSF/plasma concentration ratio Kp, CSF≥0.15.

The present invention further relates to the following Compounds 1-29,and 31-54, or their pharmaceutically acceptable salts, prodrugs, stableisotope derivatives, isomers and mixtures thereof.

Compound No. Compound Structure and Chemical Name  1.

N-(4-(4-amino-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide  2.

N-(4-(4-amino-1-(4-(hydroxymethyl)cyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide  3.

N-(4-(4-amino-1-((1s,4s)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide  4.

N-(4-(4-amino-1-((1r,4r)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide  5.

N-(4-(8-amino-3-((1r,4r)-4-(hydroxymethyl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide  6.

N-(4-(8-amino-3-(4-(hydroxymethyl)piperidin-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide  7.

N-(4-(8-amino-3-((1r,4r)-4-(aminomethyl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide  8.

N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide  9.

N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)picolinamide 10.

N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-2-methoxybenzamide 11.

N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)-2-fluorobenzyl)-2-methoxybenzamide 12.

N-(4-(4-amino-7-cyclopentylimidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 13.

(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylic acid 14.

N-(4-(4-amino-7-((1r,4r)-4-(hydroxymethyl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 15.

(1r,4r)-4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexane-1-carboxylic acid 16.

N-(4-(4-amino-7-(tetrahydro-2H-pyran-3-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 17.

N-(4-(4-amino-7-(tetrahydrofuran-3-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 18.

N-(4-(4-amino-7-((1r,4r)-4-(hydroxymethyl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-(methoxy-d₃)benzamide 19.

N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-3-methoxy-6-methylpicolinamide 20.

N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-methoxy-2-methylisonicotinamide 21.

N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide 22.

(R)-N-(4-(4-amino-7-(pyrrolidin-3-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 23.

N-(4-(8-amino-3-((1r,4r)-4-(methylcarbamoyl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide 24.

(R)-N-(4-(4-amino-7-(1-(cyclopropanecarbonyl)pyrrolidin-3-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide25.

N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-(methoxy-dz)benzamide 26.

N-(4-(4-amino-7-(1,1,1-trifluoropropan-2-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 27.

1-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-4-methylpiperidine-4-carboxylic acid 28.

methyl 4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)bicyclo[2.2.2]octane-1-carboxylate 29.

methyl 3-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)bicyclo[1.1.1]pentane-1-carboxylate 31.

3-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)bicyclo[1.1.1]pentane-1-carboxylic acid 32.

(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-1,4-dimethylcyclohexane-1-carboxylic acid 33.

methyl 4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylate 34.

4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylic acid 35.

N-(4-(8-amino-3-(4-(hydroxymethyl)bicyclo[2.2.2]octan-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide 36.

4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-N-methylbicyclo[2.2.2]octane-1-carboxamide 37.

N-(4-(8-amino-3-(4-(morpholine-4-carbonyl)bicyclo[2.2.2]octan-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide 38.

methyl 4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.1]heptane-1-carboxylate 39.

methyl (1s,4s)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-1,4-dimethylcyclohexane-1-carboxylate 40.

4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.1]heptane-1-carboxylic acid 41.

(1s,4s)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-1,4-dimethylcyclohexane-1-carboxylic acid 42.

N-(4-(8-amino-3-(1-(trifluoromethyl)cyclopropyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide 43.

N-(4-(4-amino-7-(1-(trifluoromethyl)cyclopropyl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 44.

N-(4-(8-amino-3-morpholinoimidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide 45.

N-(4-(4-amino-7-morpholinoimidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide 46.

(1s,4s)-4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f ][1,2,4]triazin-7-yl)-1,4-dimethylcyclohexane-1-carboxylic acid 47.

(1r,4r)-4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)-1,4-dimethylcyclohexane-1-carboxylic acid 48.

4-(6-amino-7-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)bicyclo[2.2.2]octane-1-carboxylic acid 49.

(1s,4s)-4-(6-amino-7-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)-8-oxo-7,8-dihydro-9H-purin-9-yl)-1,4-dimethylcyclohexane-1-carboxylicacid 50.

N-(4-(6-amino-9-((1r,4r)-4-(hydroxymethyl)cyclohexyl)-8-oxo-8,9-dihydro-7H-purin-7-yl)benzyl)-5-fluoro-2-methoxybenzamide 51.

N-(4-(6-amino-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-7-yl)benzyl)-5-fluoro-2-methoxybenzamide 52.

N-(4-(4-amino-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-1,2-dihydro-3H-imidazo[4,5-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide 53.

N-(4-(6-amino-8-oxo-9-(1,1,1-trifluoropropan-2-yl)-8,9-dihydro-7H-purin-7-yl)benzyl)-5-fluoro-2-methoxybenzamide 54.

N-(4-(6-amino-9-(4-(morpholine-4-carbonyl)bicyclo[2.2.2]octan-1-yl)-8-oxo-8,9-dihydro-7H-purin-7-yl)benzyl)-5-fluoro-2-methoxybenzamide

Compounds of the present invention effectively inhibit the activities ofBTK and its C481 mutant, preferably having an IC₅₀ of less than 100 nM,and more preferably less than 10 nM.

The present invention also relates to pharmaceutical compositionscomprising compounds of Formula (I) or their pharmaceutically acceptablesalts, stable isotope derivatives, isomers and prodrugs thereof, and oneor more pharmaceutically acceptable carriers or excipients.

The present invention further relates to a pharmaceutical compositioncomprising a compound of Formula (I) or its pharmaceutically acceptablesalt, stable isotope derivative, isomer, prodrug and a mixture thereof,and at least one additional therapeutic agent, wherein the agent may bea small molecule chemotherapeutic drug (such as anti-inflammatorysteroid drug, kinase targeting drug, apoptosis inhibitor, inflammationmodulator, cytotoxic drug, DNA damage related drug) or a macromolecularimmune and/or inflammation modulator (such as CD-20 antibody, CD19antibody, PD-1 antibody).

Compounds of Formula (I) and another therapeutic agent may be present inthe same pharmaceutical composition or in different pharmaceuticalcompositions. Compounds of Formula (I) and another agent may beadministered simultaneously or sequentially in the same or differentforms.

The present invention provides a method for treating or preventingdiseases mediated by BTK or its C481 mutant. The method comprisesadministering to a patient in need a therapeutically effective amount ofcompounds of Formula (I) or their pharmaceutically acceptable salts,stable isotope derivatives, isomers, prodrugs and mixtures thereof, orpharmaceutical compositions containing compounds of Formula (I). Thediseases include but are not limited to cancer, lymphoma, leukemia,autoimmune or inflammation diseases, such as B-cell malignancies, B-celllymphoma, diffuse large B-cell lymphoma, chronic lymphocytic leukemia,small lymphocytic lymphoma, non-Hodgkin's lymphoma (such as ABC-DLBCL),mantle cell lymphoma, follicular lymphoma, Waldenstrom'smacroglobulinemia, marginal zone lymphoma, central nervous systemlymphoma, chronic lymphocytic lymphoma, B-cell prelymphocytic leukemia,plasma cell lymphoma, multiple myeloma, various solid tumors (such aslung cancer, prostate cancer, head and neck cancer, breast cancer,ovarian cancer, uterine cancer, pancreatic cancer, colon cancer, rectalcancer, stomach cancer, esophageal cancer, brain cancer, liver cancer,kidney cancer, skin cancer, muscle cancer, epithelial cancer, bladdercancer, neuroblastoma, melanoma, bone cancer, melanoma), arthritis,multiple sclerosis, osteoporosis, inflammatory bowel disease, colitis,Crohn's disease, lupus, rheumatoid arthritis, psoriatic arthritis, lupusnephritis, Sjogren's syndrome, IgG4-related diseases, idiopathicthrombocytopenic purpura, immune thrombocytopenia, Wright syndrome,psoriasis, Behcet's disease, asthma, pemphigus, diabetes, myastheniagravis, Guillain-Barre syndrome, Graves' disease, Hashimoto'sthyroiditis, vasculitis, autoimmune vasculitis, granuloma with multiplevasculitis, autoimmune hepatitis, especially B-cell lymphoma, diffuselarge B-cell lymphoma, chronic lymphocytic leukemia, small lymphocyticlymphoma, non-Hodgkin's lymphoma (such as ABC-DLBCL), mantle celllymphoma, filtration alveolar lymphoma, Waldenstrom's macroglobulinemia,marginal zone lymphoma, central nervous system lymphoma, chroniclymphocytic lymphoma, rheumatoid arthritis, systemic lupuserythematosus, multiple sclerosis, lupus nephritis, dryness Syndrome,IgG4-related diseases, idiopathic thrombocytopenic purpura, immunethrombocytopenia, pemphigus, urticaria, etc.

According to the present invention, the pharmaceutical composition maybe in any dosage form, including but not limited to tablets, capsules,solutions, freeze-dried preparations and injectable.

The pharmaceutical formulation of the present invention may beadministered in form of a dosage unit containing a predetermined amountof active ingredient. Such a unit may contain 1 mg to 1 g, preferably 5mg to 700 mg, particularly preferably 10 mg to 500 mg of a compound ofthe present invention, depending on the disease being treated, themethod of administration, as well as age, weight, and condition of thepatients. The pharmaceutical formulation may be prepared using methodswell-known in the pharmaceutical field, for example, by formulating theactive ingredient with one or more excipients or one or more adjuvants.

The pharmaceutical formulation of the present invention is suitable foradministration by any appropriate method, such as by oral (includingoral or sublingual) or parenteral (including subcutaneous,intramuscular, intravenous, or intradermal).

The present invention also provides methods for preparing compounds. Thepreparation of compounds of the present invention may be accomplished bythe following exemplary methods and examples, but these methods andexamples should not be considered as limitation of the scope of thepresent invention in any way. Compounds of the present invention mayalso be synthesized by synthetic techniques known to those skilled inthe art or by combinations of methods known in the art and of thepresent invention. The products obtained at each step of reaction areisolated by separation techniques known in the art, including but notlimited to extraction, filtration, distillation, crystallization, andchromatographic separation. The starting materials and chemical reagentsused for syntheses may be conventionally made based on literature (forexample, SciFinder) or purchased.

Heterocyclic compounds shown in Formula (I) of the present invention canbe synthesized according to the route shown below: 1) bromination of A1with NBS to give A2; 2) substitution of A2 by a protected amine (forexample, 2,4-dimethoxybenzylamine) to give A3; 3) Mitsunobu reactionbetween A3 and E-OH to provide A4; 4) Suzuki coupling of A4 with aphenylboronic acid to give A5; 5) deprotection of A5 to give A6. Afunctional group of E can be further derivatized to afford varioustarget compounds. For example, an ester in E can be hydrolyzed by alkali(such as LiOH) to form an acid and E containing protected amine oralcohol can be deprotected to give amine or alcohol, and the amine canbe further amidated to afford amides, etc.

Heterocyclic compounds shown in Formula (I) of the present invention mayalso be synthesized according to the route shown below: 1) amideformation between B1 and E-COOH to give B2; 2) cyclization of B2 inphosphine oxychloride under heated to give B3; 3) bromination of B3 withNBS to give B4; 4) substitution of B4 by a protected amine (for example,2,4-dimethoxybenzylamine) under base catalysis to give B5; 5) Suzukicoupling of B5 with a phenylboronic acid to give B6; 6) deprotection ofB6 to give B7. Similarly, a functional group of B7 can be furtherderivatized to afford various target compounds.

Heterocyclic compounds shown in Formula (I) of the present invention mayalso be synthesized according to the route shown below: 1) amideformation of C1 with E-COOH to give C2; 2) cyclization of C2 inphosphine oxychloride under heat to give C3; 3) iodination of C3 withNIS to give C4; 4) amination of C4 to give C5; 5) Suzuki coupling of C5a phenylboronic acid to give C6. Similarly, a functional group of C6 canbe further derivatized to afford various target compounds.

Heterocyclic compounds shown in Formula (I) of the present invention mayalso be synthesized according to the route shown below: 1) substitutionof D1 (A is N or CH) by E-NH₂ under base catalysis to give D2; 2)further substitution of D2 by a protected amine (such as dibenzylamine)to give D3; 3) reduction of D3 to give D4 (for example, with a reducingagent Zn/NH₄Cl); 4) cyclization of D4 with carbonyl diimidazole ortriphosgene in the present of a base (such as NEt₃) to give D5; 5)deprotection of D5 to give D6; 6) Chan-Lam coupling of D6 with aphenylboronic acid to give D7. Similarly, a functional group of D7 canbe further derivatized to afford various target compounds.

EXAMPLES

The starting materials in the present invention were synthesizedaccording to methods known in the art or purchased from ABCR GmbH & Co.KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc.,Beijing Ouhe, etc.

The structure of a compound was determined by nuclear magnetic resonance(NMR) or mass spectrometry (MS). NMR determination used a BrukerASCEND-400 NMR spectrometer. The solvent for the determination wasdeuterated dimethyl sulfoxide (DMSO-d₆), deuterated chloroform (CDCl₃)or deuterated methanol (CD₃OD). The internal standard wastetramethylsilane (TMS) and the chemical shift was given in a unit of10⁻⁶ (ppm). MS determination used an Agilent SQD (ESI) mass spectrometer(Agilent 6120).

HPLC determination used Agilent 1260 DAD high pressure liquidchromatograph (column: Poroshell 120 EC-C18, 50×3.0 mm, 2.7 μm) orWaters Arc high pressure liquid chromatograph (column: Sunfire C18,150×4.6 mm, 5 μm).

Thin layer chromatography (TLC) used GF254 silica gel plates fromQingdao Haiyang Chemical Co., Ltd. with a thickness of 0.15 to 0.2 mm,and the separation/purification of products by thin layer chromatographyused silica plates with a thickness 0.4 to 0.5 mm.

Column chromatography generally used 200 to 300 mesh silica gel fromQingdao Haiyang Chemical Co., Ltd.

Unless otherwise specified in the examples, reactions were run in roomtemperature (20-30° C.) and under an atmosphere of argon or nitrogenusing a balloon with a volume of about 1 L.

Hydrogenation was carried out under an atmosphere of hydrogen using aballoon with a volume of about 1 L that was attached to the reactionvessel after being vacuumed and filled with hydrogen repeatedly for 3times.

The microwave reaction used a CEM Discover-SP microwave reactor.

The reaction was monitored using Agilent LCMS (1260/6120) or thin layerchromatography. The solvent eluting systems for column chromatographyand TLC included a) dichloromethane/methanol, b) petroleum ether/ethylacetate, or other systems as indicated. The ratio of the solvents wasadjusted according to the polarity of the compound, and further adjustedby addition of a small amount of TEA, or an acidic or alkaline reagentas needed. The compound purification was alternatively done usingWaters' MS-guided automated preparation system (abbreviated asprep-HPLC) with a MS detector (SQD2), eluting at a flow rate of 20mL/min in an appropriate acetonitrile/water (containing 0.1% TFA orformic acid) or acetonitrile/water (containing 0.05% of 25-28% ammoniumhydroxide) gradient (XBridge-C18, 19×150 mm, 5 μm). Some compounds wereprepared as HCl salts after prep-HPLC purification by addition of 1 NHCl to the collected fractions, followed by drying under reducedpressure.

The abbreviation DMF refers to N,N-dimethylformamide.

The abbreviation DIPEA refers to N,N-diisopropylethylamine.

The abbreviation DBU refers to 1,8-diazabicycloundec-7-ene.

The abbreviation NBS refers to N-bromosuccinimide.

The abbreviation NIS refers to N-iodosuccinimide.

Pd(dppf)Cl₂ refers to[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium.

The abbreviation HATU refers to2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate.

The abbreviation LDA refers to lithium diisopropylamide.

Example 1.N-(4-(4-amino-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 1)

Step 1. (4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)boronic acid (1b)

To a solution of 5-fluoro-2-methoxybenzoic acid 1a (340 mg, 2 mmol) inDMF (0.05 mL) and dichloromethane (10 mL) at 0° C. was added oxalylchloride (279 mg, 2.2 mmol). The mixture was gradually warmed to roomtemperature and stirred for 1 hour, then cooled to 0° C. again, followedby addition of a suspension of (4-(aminomethyl)phenyl)boronic acidhydrochloride (374 mg, 2 mmol) and DIPEA (516 mg, 4 mmol) in THE (20mL). After stirring at room temperature for 15 hours, the solvent wasremoved under reduced pressure. The residue was dissolved in ethylacetate (100 mL) and washed with saturated ammonium chloride (50 mL) andbrine (50 mL). The organic phase was dried over anhydrous sodiumsulfate, filtered and concentrated under reduced pressure. The residuewas dried to give the title compound 1b (460 mg, 76%).

MS m/z (ESI): 304 [M+1]

Step 2. 3-bromo-4-chloro-1H-pyrazolo[4,3-c]pyridine (1d)

To a solution of 4-chloro-1H-pyrazolo[4,3-c]pyridine 1c (921 mg, 6 mmol)in acetonitrile (60 mL) was added NBS (1.12 g, 6.3 mmol). After stirringfor 5 hours, the solvent was removed under reduced pressure and theresidue was dried to give the title compound 1d (2.1 g). The product wasused directly in the next step without further purification.

MS m/z (ESI): 232 [M+1]

Step 3.3-bromo-N-(2,4-dimethoxybenzyl)-1H-pyrazolo[4,3-c]pyridin-4-amine (1e)

A mixture of 1d (2.1 g, crude product), 2,4-dimethoxybenzylamine (2 g,12 mmol) and DIPEA (3.1 g, 24 mmol) in acetonitrile (50 mL) was heatedto 120° C. and stirred for 15 hours. After cooling to room temperature,the solvent was removed under reduced pressure and the residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=30/1 to 1/2) to give the title compound 1e (1.2 g, 55% over twosteps).

MS m/z (ESI): 363 [M+1]

Step 4.3-bromo-N-(2,4-dimethoxybenzyl)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine(1f)

To a mixture of 1e (363 mg, 1 mmol), tetrahydro-2H-pyran-4-ol (306 mg, 3mmol) and triphenylphosphine (524 mg, 2 mmol) in THE (5 mL) was added asolution of diisopropyl azodicarboxylate (404 mg, 2 mmol) in THE (5 mL)dropwise. After stirring for 3 hours, the solvent was removed underreduced pressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=50/1 to 1/1) to give thetitle compound 1f (220 mg, 49%).

MS m/z (ESI): 447 [M+1]

Step 5.N-(4-(4-((2,4-dimethoxybenzyl)amino)-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(1g)

A mixture of 1f (134 mg, 0.3 mmol), 1b (90 mg, 0.3 mmol), potassiumcarbonate (83 mg, 0.6 mmol) and PdCl₂(dppf) (22 mg, 0.03 mmol) in1,4-dioxane (3 mL) and water (1 mL) was heated to 120° C. under anitrogen atmosphere and stirred for 1 hour. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography (petroleumether/ethyl acetate=20/1 to 1/5) to give the title compound 1g (80 mg,42%).

MS m/z (ESI): 626 [M+1]

Step 6.N-(4-(4-amino-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(1)

Trifluoroacetic acid (5 mL) was added to a solution of 1g (80 mg, 0.13mmol) in dichloromethane (2 mL) and the resulting mixture was heated to40° C. and stirred for 2 hours. After cooling to room temperature, thesolvent was removed under reduced pressure and the residue was purifiedby prep-HPLC to give the title compound 1 (21 mg, solid, 34%).

MS m/z (ESI): 476 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.83-7.50 (m, 6H), 7.32-7.25 (m, 1H),7.24-7.11 (m, 2H), 4.84 (d, J=11.7 Hz, 1H), 4.73 (s, 2H), 4.13 (dd,J=11.4, 3.7 Hz, 2H), 4.00 (s, 3H), 3.68 (t, J=11.2 Hz, 2H), 2.37 (ddd,J=16.7, 12.5, 4.8 Hz, 2H), 1.98 (dd, J=11.2, 9.0 Hz, 2H).

Example 2.N-(4-(4-amino-1-(4-(hydroxymethyl)cyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 2)

Step 1. methyl4-(3-bromo-4-((2,4-dimethoxybenzyl)amino)-1H-pyrazolo[4,3-c]pyridin-1-yl)cyclohexane-1-carboxylate(2a)

To a mixture of 1e (726 mg, 2 mmol), methyl4-hydroxycyclohexane-1-carboxylate (948 mg, 6 mmol) andtriphenylphosphine (733 mg, 2.8 mmol) in THE (30 mL) was addeddiisopropyl azodicarboxylate (566 mg, 2.8 mmol) dropwise. After stirringfor 15 hours, the solvent was removed under reduced pressure and theresidue was purified by prep-HPLC to give the title compound 2a (130 mg,13%).

MS m/z (ESI): 503 [M+1]

Step 2. methyl4-(4-((2,4-dimethoxybenzyl)amino)-3-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)-1H-pyrazolo[4,3-c]pyridin-1-yl)cyclohexane-1-carboxylate(2b)

A mixture of 2a (130 mg, 0.26 mmol), 1b (94 mg, 0.32 mmol), potassiumcarbonate (72 mg, 0.52 mmol) and PdCl₂(dppf) (19 mg, 0.026 mmol) in1,4-dioxane (4 mL) and water (1 mL) was heated to 120° C. and stirredfor 1 hour. After cooling to room temperature, the solvent was removedunder reduced pressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=20/1 to 0/100) to give thetitle compound 2b (70 mg, 39%).

MS m/z (ESI): 682 [M+1]

Step 3.N-(4-(4-((2,4-dimethoxybenzyl)amino)-1-(4-(hydroxymethyl)cyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(2c)

To a solution of 2b (70 mg, 0.1 mmol) in THE (10 mL) at 0° C. was addedlithium aluminum hydride (76 mg, 2 mmol). After stirring for 1 hour at0° C., saturated brine (0.5 mL) was added, and the mixture was filtered.The filtrate was concentrated to dryness to afford the title compound 2c(45 mg, 69%).

MS m/z (ESI): 654 [M+1]

Step 4.N-(4-(4-amino-1-(4-(hydroxymethyl)cyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(2)

To a solution of 2c (45 mg, 0.069 mmol) in dichloromethane (1 mL) wasadded trifluoroacetic acid (3 mL). The resulting mixture was heated to50° C. and stirred for 2 hours. After cooling to room temperature, thesolvent was removed under reduced pressure and the residue was purifiedby prep-HPLC to give the title compound 2 (2.8 mg, solid, 8%).

MS m/z (ESI): 504 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.78-7.62 (m, 4H), 7.58 (d, J=8.1 Hz, 2H),7.32-7.24 (m, 1H), 7.20 (dd, J=9.1, 4.2 Hz, 1H), 6.91 (d, J=6.4 Hz, 1H),4.73 (s, 2H), 4.61-4.55 (m, 1H), 4.00 (s, 3H), 3.68 (d, J=7.3 Hz, 2H),2.21 (dd, J=21.5, 10.1 Hz, 2H), 2.00-1.75 (m, 7H).

Example 3.N-(4-(4-amino-1-((1s,4s)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 3) andN-(4-(4-amino-1-((1r,4r)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 4)

Step 1.3-bromo-N-(2,4-dimethoxybenzyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine(3a)

To a mixture of 1e (600 mg, 1.65 mmol), 1,4-dioxaspiro[4.5]decane-8-ol(521 mg, 3.3 mmol) and triphenylphosphine (864 mg, 3.3 mmol) in THE (30mL) was added diisopropyl azodicarboxylate (667 mg, 3.3 mmol) dropwise.After stirring for 15 hours, the solvent was removed under reducedpressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=50/1 to 2/1) to give thetitle compound 3a (440 mg, 53%).

MS m/z (ESI): 503 [M+1]

Step 2.N-(4-(4-((2,4-dimethoxybenzyl)amino)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(3b)

A mixture of 3a (250 mg, 0.5 mmol), 1b (151 mg, 0.5 mmol), potassiumcarbonate (138 mg, 1 mmol) and PdCl₂(dppf) (36 mg, 0.05 mmol) in1,4-dioxane (5 mL) and water (1 mL) was heated to 120° C. under anitrogen atmosphere and stirred for 1 hour. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography (petroleumether/ethyl acetate=50/1 to 1/3) to give the title compound 3b (100 mg,29%).

MS m/z (ESI): 682 [M+1]

Step 3.N-(4-(4-amino-1-(4-oxocyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(3c)

To a solution of 3b (100 mg, 0.15 mmol) in dichloromethane (1 mL) wasadded trifluoroacetic acid (4 mL). The resulting mixture was heated to50° C. and stirred for 3 hours. After cooling to room temperature, thesolvent was removed under reduced pressure to give the title compound 3c(110 mg). The product was used directly in the next step without furtherpurification.

MS m/z (ESI): 488 [M+1]

Step 4.N-(4-(4-amino-1-((1s,4s)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 3) andN-(4-(4-amino-1-((1r,4r)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 4)

To a solution of 3c (110 mg, crude product) in THE (15 mL) at 0° C. wasadded lithium aluminum hydride (56 mg, 1.47 mmol). After stirring for 1hour, the mixture was added with saturated brine (1 mL), filtered, andconcentrated to dryness. The residue was purified by prep-HPLC to givethe title compound 3 (7.3 mg, solid, 10%) and 4 (5.1 mg, solid, 7%).

N-(4-(4-amino-1-((1s,4s)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(3)

MS m/z (ESI): 490 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.73 (d, J=6.3 Hz, 1H), 7.66 (td, J=6.2, 2.8Hz, 3H), 7.58 (d, J=8.3 Hz, 2H), 7.28 (ddd, J=9.1, 7.6, 3.3 Hz, 1H),7.19 (dd, J=9.1, 4.2 Hz, 1H), 6.94 (d, J=6.4 Hz, 1H), 4.73 (s, 2H), 4.52(t, J=11.5 Hz, 1H), 3.99 (s, 3H), 3.76-3.70 (m, 1H), 2.21-2.03 (m, 6H),1.63-1.53 (m, 2H).

N-(4-(4-amino-1-((1r,4r)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(4)

MS m/z (ESI): 490 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.72-7.64 (m, 4H), 7.58 (d, J=8.2 Hz, 2H),7.28 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.20 (dd, J=9.1, 4.2 Hz, 1H), 7.02(d, J=6.5 Hz, 1H), 4.73 (s, 2H), 4.60-4.54 (m, 1H), 4.06 (s, 1H), 4.00(s, 3H), 2.49 (dd, J=13.3, 10.3 Hz, 2H), 2.01 (d, J=12.9 Hz, 2H), 1.82(t, J=13.7 Hz, 4H).

Example 4.N-(4-(4-amino-1-((1r,4r)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 5)

Step 1. methyl(1r,4r)-4-(((3-chloropyrazin-2-yl)methyl)carbamoyl)cyclohexane-1-carboxylate(5b)

To a mixture of (3-chloropyrazin-2-yl)methylamine hydrochloride 5a (360mg, 2 mmol), (1r,4r)-4-(methoxycarbonyl)cyclohexane-1-carboxylic acid(373 mg, 2 mmol) and triethylamine (606 mg, 6 mmol) in DMF (10 mL) wasadded HATU (836 mg, 2.2 mmol). After stirring for 16 hours, the reactionmixture was added with water (20 mL) and extracted with ethyl acetate(3×20 mL). The combined organic phase was dried over anhydrous sodiumsulfate, filtered, and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography (petroleum ether/ethylacetate=10/1 to 4/1) to give the title compound 5b (520 mg, 84%).

MS m/z (ESI): 312 [M+1]

Step 2. methyl(1r,4r)-4-(8-chloroimidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylate(5c)

To a solution of 5b (520 mg, 1.67 mmol) in acetonitrile (10 mL) at 0° C.was added phosphine oxychloride (2.55 g, 16.7 mmol) in portions. Theresulting mixture was heated to 100° C. and stirred for 2 hours. Aftercooling to 0° C., the solvent was removed under reduced pressure to givethe title compound 5c (420 mg, 86%). The product was used directly inthe next reaction without further purification.

MS m/z (ESI): 294 [M+1]

Step 3. methyl(1r,4r)-4-(1-bromo-8-chloroimidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylate(5d)

To a solution of 5c (420 mg, 1.43 mmol) in acetonitrile (100 mL) at 0°C. was added NBS (280 mg, 1.57 mmol). After stirring at room temperaturefor 1 hour, the mixture was quenched with water (20 mL) and extractedwith dichloromethane (2×20 mL). The combined organic phase was driedover anhydrous sodium sulfate, filtered, and concentrated under reducedpressure. The residue was purified by prep-HPLC to give the titlecompound 5d (180 mg, 34%).

MS m/z (ESI): 372 [M+1]

Step 4. methyl(1r,4r)-4-(1-bromo-8-((2,4-dimethoxybenzyl)amino)imidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylate(5e)

To a solution of 5d (160 mg, 0.43 mmol) in acetonitrile (5 mL) wereadded 2,4-dimethoxybenzylamine (49 mg, 0.47 mmol) and DIPEA (166 mg, 1.3mmol). The mixture was heated to 50° C. and stirred for 18 hours. Aftercooling to room temperature, the solvent was removed under reducedpressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=100/0 to 7/3) to give thetitle compound 5e (160 mg, 74%).

MS m/z (ESI): 503 [M+1]

Step 5. methyl(1r,4r)-4-(8-((2,4-dimethoxybenzyl)amino)-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylate(5f)

To a solution of 5e (160 mg, 0.318 mmol) in 1,4-dioxane (5 mL) wereadded 1b (191 mg, 0.636 mmol), potassium carbonate (132 mg, 0.954 mmol)and PdCl₂(dppf) (24 mg, 0.032 mmol).

The mixture was heated to 100° C. under a nitrogen atmosphere andstirred for 4 hours. After cooling to room temperature, the solvent wasremoved under reduced pressure and the residue was purified by silicagel column chromatography (petroleum ether/ethyl acetate=100/0 to 7/3)to give the title compound 5f (140 mg, 64%).

MS m/z (ESI): 682 [M+1]

Step 6.N-(4-(8-((2,4-dimethoxybenzyl)amino)-3-((1r,4r)-4-(hydroxymethyl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(5g)

To a solution of 5f (140 mg, 0.204 mmol) in THE (5 mL) at 0° C. wasadded lithium aluminum hydride (23 mg, 0.616 mmol). After stirring atroom temperature for 1 hour, it was cooled to 0° C. and to which 20%NaOH aqueous solution (0.1 mL) was added dropwise. The resulting mixturewas stirred at room temperature for 1 hour, filtered and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=100/0 to 1/1) to give thetitle compound 5g (100 mg, 75%).

MS m/z (ESI): 654 [M+1]

Step 7.N-(4-(4-amino-1-((1r,4r)-4-hydroxycyclohexyl)-1H-pyrazolo[4,3-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(5)

A mixture of 5g (80 mg, 0.122 mmol) and trifluoroacetic acid (5 mL) washeated to 80° C. and stirred for 1 hour. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by prep-HPLC to give the title compound 5 (12.0 mg,solid, 20%).

MS m/z (ESI): 504 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 8.91-8.76 (m, 1H), 7.61 (d, J=5.1, 1H),7.58-7.50 (m, 3H), 7.45 (d, J=8.1, 2H), 7.40-7.28 (m, 1H), 7.19 (dd,J=9.0, 4.3, 1H), 7.00 (d, J=4.9, 1H), 5.94 (s, 2H), 4.57 (d, J=6.0, 2H),4.51-4.34 (m, 1H), 3.90 (s, 3H), 3.27 (d, J=5.6, 1H), 3.12-3.00 (m, 2H),1.97 (d, J=11.5, 2H), 1.85 (d, J=10.7, 2H), 1.62 (d, J=14.0, 2H), 1.45(s, 2H), 1.25 (d, J=9.5, 1H).

The intermediate as shown below was synthesized according to theprocedures for the first to fourth steps in Example 4, except thattetrahydro-2H-pyran-3-carboxylic acid was used instead of(1r,4r)-4-(methoxycarbonyl)cyclohexane-1-carboxylic acid in the firststep.

Compound replacing (1r,4r)-4- MS Intermediate(methoxycarbonyl)cyclohexane- m/z No. Structure 1-carboxylic acid (ESI)19e

447

Example 5.N-(4-(8-amino-3-(4-(hydroxymethyl)piperidin-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 6)

Step 1. methyl1-(((3-chloropyrazin-2-yl)methyl)carbamoyl)piperidine-4-carboxylate (6a)

To a solution of (3-chloropyrazine-2-yl) methylamine hydrochloride 5a(1.80 g, 10 mmol) and DIPEA (1.29 g, 10 mmol) in DMF (10 mL) was addedN,N′-carbonyldiimidazole (1.62 g, 10 mmol). The mixture was stirred for3 hours and then added with methyl piperidine-4-carboxylate (1.43 g, 10mmol). The mixture was stirred for additional 18 hours and thenfiltered. The filter cake was washed with ethyl acetate (3×10 mL). Thecombined filtrates were washed with water (3×10 mL), dried overanhydrous sodium sulfate, filtered, and concentrated under reducedpressure to give the title compound 6a (3 g, 96%). The product was useddirectly in the next step without further purification.

MS m/z (ESI): 313 [M+1]

Step 2. methyl1-(8-chloroimidazo[1,5-a]pyrazin-3-yl)piperidine-4-carboxylate (6b)

To a solution of 6a (3 g, 9.6 mmol) in acetonitrile (10 mL) was addedphosphorus oxychloride (2.98 g, 19.2 mmol). The mixture was heated to100° C. and stirred for 3 hours. After cooling to room temperature, theresidue was dispersed in ethyl acetate (10 mL) and adjusted to pH=9 withsaturated sodium carbonate solution. The resulting mixture was extractedwith ethyl acetate (3×20 mL). The combined organic phase was washed withwater (3×10 mL), dried over anhydrous sodium sulfate, filtered, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (petroleum ether/ethyl acetate=100/1 to 4/1)to give the title compound 6b (260 mg, 9%).

MS m/z (ESI): 295 [M+1]

Step 3. methyl1-(1-bromo-8-chloroimidazo[1,5-a]pyrazin-3-yl)piperidine-4-carboxylate(6c)

To a solution of 6b (260 mg, 0.88 mmol) in acetonitrile (5 mL) was addedNBS (189 mg, 1.1 mmol). After the mixture was stirred for 5 hours, thesolvent was removed under reduced pressure and the residue was purifiedby silica gel column chromatography (petroleum ether/ethyl acetate=100/1to 17/3) to give the title compound 6c (200 mg, 67%).

MS m/z (ESI): 373 [M+1]

Step 4. methyl1-(1-bromo-8-((2,4-dimethoxybenzyl)amino)imidazo[1,5-a]pyrazin-3-yl)piperidine-4-carboxylate(6d)

To a solution of 6c (200 mg, 0.54 mmol) and 2,4-dimethoxybenzylamine (99mg, 0.59 mmol) in acetonitrile (5 mL) was added DIPEA (208 mg, 1.6mmol). The mixture was heated to 60° C. and stirred for 18 hours. Aftercooling to room temperature, the solvent was removed under reducedpressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=100/1 to 7/3) to give thetitle compound 6d (200 mg, 74%).

MS m/z (ESI): 504 [M+1]

Step 5. methyl1-(8-((2,4-dimethoxybenzyl)amino)-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)piperidine-4-carboxylate(6e)

To a mixture of 6d (200 mg, 0.4 mmol), 1b (145 mg, 0.48 mmol) andPdCl₂(dppf) (29 mg, 0.04 mmol) in 1,4-dioxane (10 mL) were addedpotassium carbonate (82 mg, 0.6 mmol) and water (2 mL). The mixture washeated to 100° C. under a nitrogen atmosphere and stirred for 3 hours.After cooling to room temperature, the mixture was extracted with ethylacetate (3×10 mL). The combined organic phase was washed with water(3×10 mL), dried over anhydrous sodium sulfate, filtered, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (petroleum ether/ethyl acetate=100/1 to 4/1)to give the title compound 6e (170 mg, 63%).

MS m/z (ESI): 683 [M+1]

Step 6.N-(4-(8-((2,4-dimethoxybenzyl)amino)-3-(4-(hydroxymethyl)piperidin-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(6f)

To a solution of 6e (90 mg, 0.13 mmol) in THE (3 mL) at 0° C. was addedlithium aluminum hydride (10 mg, 0.26 mmol). After the mixture wasgradually warmed to room temperature and stirred for 3 hours, ethylacetate (1 mL) and water (1 mL) were added. After stirring for 10minutes, the mixture was extracted with ethyl acetate (3×5 mL). Thecombined organic phase was washed with water (3×5 mL), dried overanhydrous sodium sulfate, filtered, and concentrated under reducedpressure to give the title compound 6f (100 mg). The product was useddirectly in the next step without further purification.

MS m/z (ESI): 655 [M+1]

Step 7.N-(4-(8-amino-3-(4-(hydroxymethyl)piperidin-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(6)

To a solution of 6f (100 mg, 0.15 mmol) in dichloromethane (5 mL) wasadded trifluoroacetic acid (2 mL). The mixture was heated to 50° C. andstirred for 18 hours. After cooling to room temperature, the solvent wasremoved under reduced pressure, and the residue was dissolved inmethanol (2 mL) and to which potassium carbonate (138 mg) was added. Themixture was stirred for 3 hours, filtered, and concentrated underreduced pressure. The residue was purified by prep-HPLC to give thetitle compound 6 (27.6 mg, solid, 42%).

MS m/z (ESI): 505 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 8.84 (t, J=6.1 Hz, 1H), 7.62-7.48 (m, 3H),7.44 (d, J=8.2 Hz, 2H), 7.37-7.31 (m, 1H), 7.22-7.14 (m, 2H), 6.95 (d,J=4.9 Hz, 1H), 5.99-5.87 (m, 2H), 4.61-4.48 (m, 3H), 3.90 (s, 3H),3.42-3.33 (m, 4H), 2.89-2.79 (m, 2H), 1.82-1.74 (m, 2H), 1.59-1.52 (m,1H), 1.44-1.34 (m, 2H).

The intermediate as shown below was synthesized according to theprocedures for the first to the fifth steps in Example 5, except thatmethyl 4-methylpiperidine-4-carboxylate was used instead of methylpiperidine-4-carboxylate in the first step.

MS Intermediate Compound replacing methyl m/z No. Structurepiperidine-4-carboxylate (ESI) 27a

697

Compound 44 was synthesized according to the procedure in Example 5(without the sixth step), except that morpholine was used instead ofmethyl piperidine-4-carboxylate in the first step.

MS Compound Compound replacing methyl m/z No. Structurepiperidine-4-carboxylate (ESI) 44

477

¹H NMR data of Compound 44 is shown below:

Compound ¹H NMR N-(4-(8-amino-3- ¹H NMR (400 MHz, DMSO-d₆) δ 8.85 (t, J= morpholinoimidazo[1,5- 6.1 Hz, 1H), 8.14 (s, 0.5H), 7.59 (d, J = 8.2a]pyrazin-1-yl)benzyl)- Hz, 2H), 7.52 (m, 1H), 7.46 (d, J = 8.2 Hz,5-fluoro-2- 2H), 7.40-7.30 (m, 2H), 7.19(m, 1H), 6.96 (d,methoxybenzamide (44) J = 5.2 Hz, 1H), 6.54 (brs, 2.5 H), 4.57 (d, J =6.1 Hz, 2H), 3.90 (s, 3H), 3.84-3.75 (m, 4H), 3.18-3.10 (m, 4H).

Example 6.N-(4-(8-amino-3-((1r,4r)-4-(aminomethyl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 7)

Step 1.N-(4-(3-((1r,4r)-4-(bromomethyl)cyclohexyl)-8-((2,4-dimethoxybenzyl)amino)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(7a)

To a slurry of 5g (160 mg, 0.25 mmol) in dichloromethane (10 mL) wereadded triphenylphosphine (96 mg, 0.37 mmol) and carbon tetrabromide (122mg, 0.37 mmol) in sequence under a nitrogen atmosphere. The mixture washeated to reflux and stirred for 18 hours. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography (petroleumether/ethyl acetate=4/1 to 1/1) to give the title compound 7a (170 mg,97%).

MS m/z (ESI): 716 [M+1]

Step 2.N-(4-(3-((1r,4r)-4-(aminomethyl)cyclohexyl)-8-((2,4-dimethoxybenzyl)amino)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(7b)

A mixture of 7a (110 mg, 0.16 mmol) and a solution of ammonia in THE (4M, 5 mL) in a sealed tube was heated to 100° C. and stirred for 48hours. After cooling to room temperature, the solvent was removed underreduced pressure and the residue was purified by silica gel columnchromatography (dichloromethane/methanol=100/0=50/1) to give the titlecompound 7b (60 mg, 58%).

MS m/z (ESI): 653 [M+1]

Step 3.N-(4-(8-amino-3-((1r,4r)-4-(aminomethyl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(7)

A mixture of 7b (60 mg, 0.092 mmol) and trifluoroacetic acid (10 mL) washeated to 80° C. and stirred for 18 hours. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by reversed phase preparative high performanceliquid chromatography to give the title compound 7 (4.5 mg, solid, 10%).

MS m/z (ESI): 503 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.73-7.41 (m, 6H), 7.37-7.15 (m, 2H), 7.00 (s,1H), 4.70 (s, 2H), 3.97 (s, 3H), 3.24-3.11 (m, 1H), 2.87 (d, J=6.1, 2H),2.20-2.00 (m, 5H), 1.92-1.66 (m, 4H).

Example 7.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 8)

Step 1. (3,4-dihydro-2H-pyran-2-yl)methyl acetate (8b)

To a solution of (3,4-dihydro-2H-pyran-2-yl)-methanol 8a (12.5 g, 110mmol) in dichloromethane (50 mL) were added triethylamine (16.7 g, 165mmol) and acetic anhydride (16.8 g, 165 mmol). After stirring for 15hours, the mixture was diluted with dichloromethane (200 mL) and washedwith saturated ammonium chloride solution (50 mL), water (50 mL) andsaturated brine (50 mL) in sequence. The solvent was removed from theorganic phase under reduced pressure and the residue was purified bysilica gel column chromatography (petroleum ether/ethyl acetate=30/1 to1/2) to give the title compound 8b (15 g, 87%).

¹H NMR (400 MHz, CDCl₃) δ 6.38 (dd, J=4.5, 1.7 Hz, 1H), 4.71 (dddd,J=6.2, 4.9, 2.6, 1.2 Hz, 1H), 4.17 (qd, J=11.6, 5.2 Hz, 2H), 4.11-3.99(m, 1H), 2.14-2.06 (m, 4H), 2.01 (dddd, J=15.7, 11.0, 2.9, 1.6 Hz, 1H),1.89-1.82 (m, 1H), 1.69 (dtd, J=13.5, 10.3, 6.0 Hz, 1H).

Step 2. (5-formyl-3,4-dihydro-2H-pyran-2-yl)methyl acetate (8c)

To DMF (80 mL) was added phosphine oxychloride (25.5 g, 166 mmol). Theresulting mixture was stirred for 30 minutes, cooled to 0° C. and addedwith 8b (13 g, 83 mmol). After warming to room temperature, the mixturewas stirred for 15 hours, then added with saturated sodium bicarbonatesolution (50 mL) and stirred for additional 15 hours. The mixture wasextracted with ethyl acetate (3×200 mL). The combined organic phase wasdried over anhydrous sodium sulfate, filtered, and concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=50/1 to 3/1) to give thetitle compound 8c (8 g, 52%).

¹H NMR (400 MHz, CDCl₃) δ 9.27 (s, 1H), 7.33 (d, J=9.4 Hz, 1H),4.34-4.20 (m, 3H), 2.43 (ddd, J=17.0, 5.5, 3.2 Hz, 1H), 2.25-2.14 (m,1H), 2.12 (s, 3H), 2.04-1.94 (m, 1H), 1.76

-   -   1.62 (m, 1H).

Step 3. 2-(acetoxymethyl)-3,4-dihydro-2H-pyran-5-carboxylic acid (8d)

To a mixture of 8c (1.5 g, 8.1 mmol), sodium phosphate monobasic (2.43g, 20.3 mmol), acetonitrile (16 mL), n-butanol (16 mL) and water (8 mL)was added hydrogen peroxide (30%, 3.67 mL, 32.4 mmol). After stirringfor 30 minutes, to the mixture was added sodium chlorite (3.66 g, 40.5mmol) and stirring was continued for additional 15 hours. The reactionmixture was diluted with saturated brine (80 mL) and extracted withethyl acetate (2×100 mL). The combined organic phase was dried overanhydrous sodium sulfate, filtrated, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(petroleum ether/ethyl acetate=50/1 to 3/1) to afford the title compound8d (1.03 g, 63%).

MS m/z (ESI): 201 [M+1]

Step 4. 6-(acetoxymethyl)tetrahydro-2H-pyran-3-carboxylic acid (8e)

A mixture of 8d (1 g, 5 mmol) and palladium on carbon (10%, 0.7 g) inethyl acetate (80 mL) was stirred for 48 hours under a hydrogenatmosphere. The mixture was filtered and concentrated to dryness to givethe title compound 8e (1.05 g, 100%). The product was used directly inthe next step without further purification.

MS m/z (ESI): 203 [M+1]

Step 5.(5-(((3-chloropyrazin-2-yl)methyl)carbamoyl)tetrahydro-2H-pyran-2-yl)methylacetate (8)

To a mixture of 5a (890 mg, 5.0 mmol), 8e (1.05 g, crude product) andDIPEA (1.28 g, 9.9 mmol) in DMF (15 mL) was added HATU (2.26 g, 5.9mmol). After stirring for 30 minutes, the mixture was purified byreversed phase preparative high performance liquid chromatography togive the title compound 8f (1.5 g, 72%).

MS m/z (ESI): 328 [M+1]

Step 6.(5-(8-chloroimidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methylacetate (8g)

A mixture of 8f (229 mg, 0.7 mmol) and phosphine oxychloride (215 mg,1.4 mmol) in acetonitrile (20 mL) was heated to 80° C. and stirred for 6hours. After cooling to 0° C., saturated sodium bicarbonate solution (30mL) was added, and the mixture was extracted with ethyl acetate (2×50mL). The combined organic phase was dried over anhydrous sodium sulfate,filtered, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=50/1 to 2/1) to give the title compound 8g (150 mg, 69%).

MS m/z (ESI): 310 [M+1]

Step 7.(5-(1-bromo-8-chloroimidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methylacetate (8h)

To a solution of 8g (150 mg, 0.48 mmol) in acetonitrile (20 mL) at 0° C.was added NBS (112 mg, 0.63 mmol). The mixture was allowed to warm toroom temperature and stirred for 30 minutes. The solvent was removedunder reduced pressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=50/1 to 2/1) to give thetitle compound 8h (170 mg, 89%).

MS m/z (ESI): 388 [M+1]

Step 8.(5-(8-amino-1-bromoimidazo[1,5-a]pyrazin-3-yl)tetrahydro-2H-pyran-2-yl)methylacetate (8i)

A mixture of 8h (150 mg, 0.386 mmol) and ammonium hydroxide (10 mL) inacetonitrile (10 mL) was heated to 80° C. and stirred for 4.5 hours in asealed tube. After cooling to room temperature, the solvent was removedunder reduced pressure to give the title compound 8i (145 mg). Theproduct was used directly in the next step without further purification

MS m/z (ESI): 369 [M+1]

Step 9.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(8)

A mixture of 8i (145 mg, 0.39 mmol), 1b (174 mg, 0.58 mmol), potassiumcarbonate (106 mg, 0.77 mmol), PdCl₂(dppf) (29 mg, 0.04 mmol),1,4-dioxane (5 mL) and water (1 mL) was heated to 120° C. under anitrogen atmosphere and stirred for 2 hours. After cooling to roomtemperature, it was diluted with saturated brine (3 mL) and extractedwith ethyl acetate (3×50 mL). The combined organic phase was dried overanhydrous sodium sulfate, filtered, and concentrated under reducedpressure. The residue was purified by prep-HPLC to give the titlecompound 8 (32 mg, solid, 16%).

MS m/z (ESI): 506 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.62 (dd, J=9.1, 3.1 Hz, 3H), 7.56-7.47 (m,3H), 7.29-7.20 (m, 1H), 7.17 (dd, J=9.1, 4.2 Hz, 1H), 6.99 (d, J=5.1 Hz,1H), 4.70 (s, 2H), 4.29 (dd, J=11.8, 3.0 Hz, 1H), 3.97 (s, 3H), 3.90(dd, J=11.8, 3.5 Hz, 1H), 3.64 (dd, J=9.6, 4.2 Hz, 2H), 3.56 (t, J=7.1Hz, 1H), 3.38 (s, 1H), 2.29 (dd, J=13.6, 4.2 Hz, 1H), 2.14-2.03 (m, 1H),1.96-1.86 (m, 1H), 1.67-1.59 (m, 1H).

Example 8.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)picolinamide(Compound 9)

Step 1. (4-(picolinamidomethyl)phenyl)boronic acid (9b)

To a mixture of picolinic acid 9a (240 mg, 1.95 mmol),(4-(aminomethyl)phenyl) borate hydrochloride (280 mg, 1.5 mmol), DIPEA(387 mg, 3 mmol) and DMF (4 mL) was added HATU (855 mg, 2.25 mmol).After stirring for 1 hour, the mixture was purified by prep-HPLC to givethe title compound 9b (295 mg, 77%).

MS m/z (ESI): 257 [M+1]

Step 2.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)picolinamide(9)

A mixture of 8i (125 mg, 0.25 mmol), 9b (128 mg, 0.5 mmol), PdCl₂(dppf)(18 mg, 0.025 mmol), potassium carbonate (69 mg, 0.5 mmol), 1,4-dioxane(4 mL) and water (1 mL) was heated to 120° C. under a nitrogenatmosphere and stirred for 1 hour. After cooling to room temperature,the solvent was removed under reduced pressure and the residue waspurified by prep-HPLC to give the title compound 9 (12 mg, solid, 11%).

MS m/z (ESI): 459 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 8.67 (d, J=4.1 Hz, 1H), 8.15 (d, J=7.8 Hz,1H), 7.99 (td, J=7.7, 1.7 Hz, 1H), 7.80-7.31 (m, 6H), 7.01 (d, J=5.2 Hz,1H), 4.74 (s, 2H), 4.31 (dd, J=11.8, 2.3 Hz, 1H), 3.92 (dd, J=11.8, 3.6Hz, 1H), 3.71-3.54 (m, 3H), 3.40 (dd, J=7.5, 3.4 Hz, 1H), 2.36-2.26 (m,1H), 2.11 (ddd, J=15.3, 9.7, 4.5 Hz, 1H), 1.99-1.88 (m, 1H), 1.69-1.60(m, 1H).

Example 9.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-2-methoxybenzamide(Compound 10)

Step 1. (4-((2-methoxybenzamido)methyl)phenyl)boronic acid (10b)

To a mixture of 2-methoxybenzoic acid (296 mg, 1.95 mmol),(4-(aminomethyl)phenyl)boronic acid hydrochloride (280 mg, 1.5 mmol),DIPEA (387 mg, 3 mmol) and DMF (4 mL) was added HATU (855 mg, 2.25mmol). After stirring for 1 hour, the mixture was purified by prep-HPLCto give the title compound 10b (270 mg, 63%).

MS m/z (ESI): 286 [M+1]

Step 2.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-2-methoxybenzamide(9)

A mixture of 8i (125 mg, 0.25 mmol), 10b (143 mg, 0.5 mmol), potassiumcarbonate (69 mg, 0.5 mmol), PdCl₂(dppf) (18 mg, 0.025 mmol),1,4-dioxane (4 mL) and water (1 mL) was heated to 120° C. under anitrogen atmosphere and stirred for 1 hour. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography(dichloromethane/methanol=50/1 to 20/1) to give the title compound 10(73 mg, solid, 60%).

MS m/z (ESI): 488 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.93 (dd, J=7.7, 1.8 Hz, 1H), 7.64 (d, J=8.2Hz, 2H), 7.60-7.49 (m, 4H), 7.18 (d, J=8.2 Hz, 1H), 7.12-7.06 (m, 1H),7.01 (d, J=5.2 Hz, 1H), 4.73 (s, 2H), 4.32 (dd, J=11.7, 2.4 Hz, 1H),4.04-3.95 (m, 3H), 3.93 (dd, J=11.8, 3.6 Hz, 1H), 3.72-3.62 (m, 2H),3.57 (dd, J=14.2, 6.9 Hz, 1H), 3.40 (dd, J=8.4, 4.3 Hz, 1H), 2.35-2.27(m, 1H), 2.17-2.06 (m, 1H), 1.99-1.89 (m, 1H), 1.69-1.61 (m, 1H).

Example 10.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)-2-fluorobenzyl)-2-methoxybenzamide(Compound 11)

Step 1. 4-bromo-2-fluorobenzaldehyde oxime (11 b)

To a solution of 4-bromo-2-fluorobenzaldehyde 11a (4.06 g, 20 mmol) inethanol (50 mL) was added hydroxylamine hydrochloride (1.53 g, 22 mmol).After stirring for 2 hours, the solvent was removed under reducedpressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=20/1 to 10/1) to give thetitle compound 11b (2.18 g, 50%).

MS m/z (ESI): 218 [M+1]

Step 2. (3-fluoro-4-((hydroxyimino)methyl)phenyl)boronic acid (11c)

To a solution of 11b (2.18 g, 10 mmol) and triisopropyl borate (3.76 g,20 mmol) in THE (50 mL) at −78° C. was added n-butyl lithium (2.5 M, 12mL, 30 mmol). After stirring for 2 hours at −78° C., water (2 mL) wasadded, and the resulting mixture was warmed to room temperature andstirred for 30 minutes. The mixture was adjusted to pH=5-6 with 1 Nhydrochloric acid and filtered. The filter cake was dried to give thetitle compound 11c (3.78 g). The product was used directly in the nextstep without further purification.

MS m/z (ESI): 184 [M+1]

Step 3. (4-(aminomethyl)-3-fluorophenyl)boronic acid (11d)

A mixture of 11c (3.7 g, 10 mmol) and palladium on carbon (10%, 500 mg)in methanol (80 mL) was stirred under a hydrogen atmosphere for 4 hours.The mixture was filtered and concentrated to dryness under reducedpressure to give the title compound 11d (2.9 g). The product was useddirectly in the next step without further purification.

MS m/z (ESI): 170 [M+1]

Step 4. (3-fluoro-4-((2-methoxybenzamido)methyl)phenyl)boronic acid(11e)

To a mixture of 2-methoxybenzoic acid (2.28 g, 15 mmol), 11d (1.69 g, 10mmol), DIPEA (3.9 g, 30 mmol) and DMF (30 mL) was added HATU (5.7 g, 15mmol). After stirring for 3 hours, the mixture was purified by prep-HPLCto give the title compound Ile (720 mg, 24%).

MS m/z (ESI): 304 [M+1]

Step 5.N-(4-(8-amino-3-(6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)-2-fluorobenzyl)-2-methoxybenzamide(11)

A mixture of 8i (125 mg, 0.25 mmol), Ile (151 mg, 0.5 mmol), potassiumcarbonate (69 mg, 0.5 mmol), PdCl₂(dppf) (18 mg, 0.025 mmol),1,4-dioxane (4 mL) and water (1 mL) was heated to 120° C. under anitrogen atmosphere and stirred for 1 hour. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography(dichloromethane/methanol=50/1 to 20/1) to give the title compound 11(55.4 mg, solid, 43%).

MS m/z (ESI): 506 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.91 (dd, J=7.8, 1.8 Hz, 1H), 7.59-7.48 (m,3H), 7.43 (ddd, J=8.2, 5.4, 1.6 Hz, 2H), 7.16 (d, J=8.2 Hz, 1H), 7.06(td, J=7.7, 0.9 Hz, 1H), 7.01 (d, J=5.1 Hz, 1H), 4.74 (s, 2H), 4.29 (dd,J=11.8, 1.9 Hz, 1H), 3.98 (d, J=5.7 Hz, 3H), 3.90 (dd, J=11.8, 3.5 Hz,1H), 3.66-3.60 (m, 2H), 3.56 (t, J=7.1 Hz, 1H), 3.38 (t, J=4.0 Hz, 1H),2.33-2.26 (m, 1H), 2.14-2.05 (m, 1H), 1.95 (s, 1H), 1.62 (dd, J=9.6, 4.2Hz, 1H).

Example 11.N-(4-(4-amino-7-cyclopentylimidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 12)

Step 1. ethyl 3-(1,3-dioxoisoindolin-2-yl)-2-hydroxypropanoate (12b)

To a solution of ethyl oxirane-2-carboxylate 12a (30 g, 259 mmol) in DMF(300 mL) were added phthalimide (34.2 g, 233 mmol) and potassiumphthalimide (9.6 g, 52 mmol) in sequence. The mixture was heated to 90°C. and stirred for 8 hours, then cooled to room temperature, dilutedwith water (600 mL), and extracted with ethyl acetate (3×600 mL). Thecombined organic phase was washed with water (3×600 mL), dried overanhydrous sodium sulfate, filtered, and concentrated under reducedpressure to give the title compound 12b (57.5 g, 84%). The product wasused directly in the next step without further purification.

MS m/z (ESI): 264 [M+1]

¹H NMR (400 MHz, CDCl₃) δ 7.87-7.85 (m, 2H), 7.80-7.69 (m, 2H), 4.50 (t,J=5.9 Hz, 1H), 4.32-4.18 (m, 2H), 4.09-3.98 (m, 2H), 3.03 (s, 1H),1.30-1.22 (m, 3H).

Step 2. ethyl 3-(1,3-dioxoisoindolin-2-yl)-2-oxopropanoate (12c)

To solution of 12b (57.5 g, 218 mmol) in acetonitrile (500 mL) at 0° C.was added 2-iodoxybenzoic acid (91.7 g, 328 mmol) in portions. Themixture was heated to 90° C. and stirred for 18 hours. After cooling toroom temperature, it was filtered. The filtrate was concentrated to avolume of about 150 mL and the newly formed precipitate was filteredout. The new filtrate was concentrated to dryness, added withdichloromethane (250 mL) and filtered. The newly obtained filtrate wasconcentrated to dryness under reduced pressure and the residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=100/0 to 1/1) to give the title compound 12c (40 g, 70%).

MS m/z (ESI): 262 [M+1]

¹H NMR (400 MHz, CDCl₃) δ 7.94-7.85 (m, 2H), 7.80-7.74 (m, 2H), 4.99 (s,2H), 4.41 (q, J=7.1 Hz, 2H), 1.41 (t, J=7.1 Hz, 3H).

Step 3.2-((5-oxo-3-thioxo-2,3,4,5-tetrahydro-1,2,4-triazin-6-yl)methyl)isoindoline-1,3-dione(12d)

To solution of 12c (40 g, 153 mmol) in acetic acid (300 mL) was addedhydrazinocarbothioformamide (14 g, 153 mmol). The mixture was heated to120° C. and stirred for 16 hours. After cooling to room temperature, theprecipitate was collected by filtration, and washed with acetic acid(2×50 mL) and petroleum ether (200 mL) to give the title compound 12d(16 g, 36%).

MS m/z (ESI): 289 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 13.38 (s, 1H), 13.29 (s, 1H), 7.96-7.85 (m,4H), 4.68 (s, 2H).

Step 4.2-((5-oxo-4,5-dihydro-1,2,4-triazin-6-yl)methyl)isoindoline-1,3-dione(12e)

To a mixture of 12d (16 g, 56 mmol) in ethanol (6 mL) was added Raneynickel (8 g, 50% wt). The mixture was heated to 85° C. and stirred for72 hours under a hydrogen atmosphere. The reaction was not complete. Thereaction mixture was cooled to room temperature and added withadditional Raney nickel (8 g, 50% wt). The mixture was heated to 85° C.and stirred for 6 hours under a hydrogen atmosphere. The hydrogenatmosphere was replaced with a nitrogen atmosphere, and the reactionmixture was quickly filtered while it was hot. The filter cake wasdissolved in ethanol (200 mL), refluxed for 1 hour and filtered (thisoperation was repeated 4 times). The combined filtrate was concentratedto dryness under reduced pressure to give the title compound 12e (11.56g, 81%).

MS m/z (ESI): 127 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 8.30 (s, 1H), 7.78 (s, 3H), 3.87 (s, 2H).

Step 5. 6-(aminomethyl)-1,2,4-triazin-5(4H)-one (12f)

To a mixture of 12e (11.56 g, 45 mmol) in ethanol (200 mL) was addedhydrazine hydrate (22.5 g, 450 mmol) dropwise. The resulting solutionwas stirred for 16 hours and then slurried with ethanol (300 mL). Afterfiltration, the solvent was removed from the filtrate under reducedpressure and the residue was purified by silica gel columnchromatography (dichloromethane/methanol (containing 10% water)=2/3 to7/3) to give the title compound 12f (700 mg). The filter cake wasslurried with water (400 mL) and filtered. The solvent was removed fromthe filtrate under reduced pressure and the residue was purified bysilica gel column chromatography (dichloromethane/methanol (containing10% water)=2/3 to 7/3) to give the title compound 12f (3.35 g). The twoportions of the product were combined to give the title compound 12f(4.05 g, 71%).

MS m/z (ESI): 127 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 8.30 (s, 1H), 7.78 (s, 3H), 3.87 (s, 2H)

Step 6.N-((5-oxo-4,5-dihydro-1,2,4-triazin-6-yl)methyl)cyclopentanecarboxamide(12g)

To a mixture of 12f (600 mg, 4.76 mmol), triethylamine (721 mg, 7.14mmol) and cyclopentanecarboxylic acid (597 mg, 5.23 mmol) indichloromethane (20 mL) was added HATU (2.72 g, 7.14 mmol). The mixturewas stirred for 18 hours, and the solvent was removed under reducedpressure. The residue was purified by silica gel column chromatography(petroleum ether/ethyl acetate=100/1 to 1/100) to give the titlecompound 12g (400 mg, 38%).

MS m/z (ESI): 223 [M+1]

Step 7. 7-cyclopentylimidazo[5,1-f][1,2,4]triazin-4(3H)-one (12h)

To a mixture of 12g (400 mg, 1.8 mmol) in acetonitrile (10 mL) was addedphosphine oxychloride (551 mg, 3.6 mmol). The mixture was heated to 100°C. and stirred for 1 hour. After cooling to room temperature, thesolvent was removed under reduced pressure and the residue was purifiedby silica gel column chromatography (petroleum ether/ethyl acetate=100/1to 1/100) to give the title compound 12h (210 mg, 54%).

MS m/z (ESI): 205 [M+1]

Step 8. 7-cyclopentyl-5-iodoimidazo[5,1-f][1,2,4]triazin-4(3H)-one (12i)

To a solution of 12h (210 mg, 1.03 mmol) in DMF (2 mL) was added NIS(242 mg, 1.08 mmol). After stirring for 18 hours, the mixture wasquenched with water (5 mL) and extracted with ethyl acetate (3×5 mL).The combined organic phase was washed with water (3×5 mL), dried overanhydrous sodium sulfate, filtered, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(petroleum ether/ethyl acetate=100/1 to 1/1) to give the title compound12i (110 mg, 31%).

MS m/z (ESI): 331 [M+1]

Step 9. 7-cyclopentyl-5-iodoimidazo[5,1-f][1,2,4]triazin-4-amine (12j)

To a solution of 1,2,4-triazole (188 mg, 2.73 mmol) in pyridine (1 mL)was added phosphine oxychloride (139 mg, 0.91 mmol). After stirring for15 minutes, a solution of 12i (110 mg, 0.33 mmol) in pyridine (1 mL) wasadded and stirring was continued for 2 hours. The mixture was cooled to0° C., and a solution of ammonia in isopropanol (2 M, 0.3 mL) was added.After stirring for 18 hours, the mixture was filtered, and the filtercake was washed with dichloromethane (5 mL). The combined filtrate wasconcentrated under reduced pressure and the residue was purified bysilica gel column chromatography (petroleum ether/ethyl acetate=100/1 to1/1) to give the title compound 12j (60 mg, 70%).

MS m/z (ESI): 330 [M+1]

Step 10.N-(4-(4-amino-7-cyclopentylimidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(12)

A mixture of 12j (40 mg, 0.12 mmol), 1b (45 mg, 0.15 mmol), potassiumcarbonate (26 mg, 0.18 mmol), PdCl₂(dppf) (9 mg, 0.012 mmol) and1,4-dioxane (2 mL) was heated to 120° C. under a nitrogen atmosphere andstirred for 18 hours. After cooling to room temperature, the mixture wasfiltered and concentrated under reduced pressure. The residue waspurified by preparative silica gel thin-layer chromatography (ethylacetate as the developing solvent) to give the title compound 12 (5.1mg, solid, 9%).

MS m/z (ESI): 461 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.73 (s, 1H), 7.57-7.49 (m, 3H), 7.44 (d,J=8.2 Hz, 2H), 7.18-7.13 (m, 1H), 7.09-7.05 (m, 1H), 4.60 (s, 2H), 3.87(s, 3H), 3.66-3.61 (m, 1H), 2.07-2.00 (m, 2H), 1.92-1.78 (m, 4H),1.68-1.61 (m, 2H).

The compounds or intermediates as shown below were synthesized accordingto the procedures in Example 11, except that a different carboxylic acidwas used instead of cyclopentanecarboxylic acid in the sixth step.

Compound/ Compound replacing MS Intermediate cyclopentanecarboxylic m/zNo. Structure acid (ESI) 14a

533 16

477 17

463

The NMR data of compounds 16 and 17 are shown below:

Compound ¹H NMR N-(4-(4-amino-7- ¹H NMR (400 MHz, CD₃OD) δ 7.88 (s, 1H),7.67-7.61 (m, (tetrahydro-2H-pyran-3- 3H), 7.59-7.54 (m, 2H), 7.30-7.25(m, 1H), 7.21-7.17 yl)imidazo[5,1- (m, 1H), 4.72 (s, 2H), 4.13 (dd, J =11.5, 2.7 Hz, 1H), 4.04- f][1,2,4]triazin-5- 3.97 (m, 4H), 3.79-3.73 (m,1H), 3.71-3.65 (m, 1H), 3.59- yl)benzyl)-5-fluoro-2- 3.52 (m, 1H),2.19-2.07 (m, 2H), 1.86-1.77 (m, 2H). methoxybenzamide (16)N-(4-(4-amino-7- ¹H NMR (400 MHz, CD₃OD) δ 8.09 (s, 1H), 7.86 (s, 1H),(tetrahydrofuran-3- 7.65-7.60 (m, 2H), 7.58-7.49 (m, 2H), 7.28-7.23 (m,yl)imidazo[5,1- 1H), 7.19-7.15 (m, 1H), 4.69 (s, 2H), 4.23 (t, J = 7.8Hz, f][1,2,4]triazin-5- 1H), 4.13-4.05 (m, 2H), 4.03-3.91 (m, 4H),2.46-2.42 yl)benzyl)-5-fluoro-2- (m, 1H), 1.32-1.26 (m, 2H).methoxybenzamide (17)

The intermediate as shown below was synthesized according to theprocedures for the sixth to ninth step in Example 11, except that(1r,4r)-4-(methoxycarbonyl)cyclohexane-1-carboxylic acid was usedinstead of cyclopentanecarboxylic acid in the sixth step.

Compound/ MS Intermediate Compound replacing m/z No. Structurecyclopentanecarboxylic acid (ESI) 18e

402

Example 12.(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylicacid (Compound 13)

Step 1. methyl(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylate(13a)

A mixture of 5f (100 mg, 0.147 mmol) and trifluoroacetic acid (10 mL)was heated to 80° C. and stirred for 2 hours. After cooling to roomtemperature, the solvent was removed under reduced pressure to give thetitle compound 13a (100 mg). The product was used directly in the nextstep without further purification.

MS m/z (ESI): 532 [M+1]

Step 2.(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)cyclohexane-1-carboxylicacid (13)

To a solution of 13a (100 mg, crude product) in methanol (5 mL) wasadded lithium hydroxide (11 mg, 0.45 mmol). After stirring for 1 hour,the solvent was removed under reduced pressure and the residue waspurified by prep-HPLC to give the title compound 13 (26.5 mg, solid,35%).

MS m/z (ESI): 518 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 12.14 (s, 1H), 8.85 (s, 1H), 7.66 (d, J=5.0,1H), 7.59-7.50 (m, 3H), 7.46 (d, J=8.1, 2H), 7.38-7.31 (m, 1H), 7.19(dd, J=9.1, 4.3, 1H), 7.02 (d, J=4.9, 1H), 5.96 (s, 2H), 4.58 (d, J=6.0,2H), 3.91 (s, 3H), 3.12 (t, J=11.4, 1H), 2.36-2.26 (m, 1H), 2.01 (t,J=10.0, 4H), 1.74-1.50 (m, 4H).

Example 13.N-(4-(4-amino-7-((1r,4r)-4-(hydroxymethyl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 14)

To a solution of 14a (40 mg, 0.075 mmol) in THE (3 mL) at 0° C. wasadded lithium aluminum hydride (6 mg, 0.15 mmol) under a nitrogenatmosphere. The mixture was stirred at room temperature for 1 hour,quenched with water (0.2 mL) and filtered. The solvent was removed fromthe filtrate under reduced pressure and the residue was purified byprep-HPLC to give the title compound 14 (15.7 mg, solid, 42%).

MS m/z (ESI): 505 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.74 (s, 1H), 7.55-7.50 (m, 3H), 7.46-7.42 (m,2H), 7.17-7.13 (m, 1H), 7.09-7.05 (m, 1H), 4.60 (s, 2H), 3.87 (s, 3H),3.33 (d, J=6.3 Hz, 2H), 2.00-1.94 (m, 2H), 1.90-1.84 (m, 2H), 1.79-1.67(m, 3H), 1.14-1.01 (m, 3H).

Example 14.(1r,4r)-4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexane-1-carboxylicacid (Compound 15)

To a mixture of 14a (40 mg, 0.075 mmol), THE (3 mL) and methanol (3 mL)was added lithium hydroxide monohydrate (13 mg, 0.3 mmol). Afterstirring for 3 hours, the solvent was removed under reduced pressure andthe residue was purified by prep-HPLC to give the title compound 15(23.1 mg, solid, 59%).

MS m/z (ESI): 519 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.86 (s, 1H), 7.68-7.61 (m, 3H), 7.59-7.54 (m,2H), 7.31-7.25 (m, 1H), 7.21-7.17 (m, 1H), 4.72 (s, 2H), 3.99 (s, 3H),3.44-3.38 (m, 1H), 2.45-2.38 (m, 1H), 2.20-2.09 (m, 4H), 1.90-1.80 (m,2H), 1.70-1.61 (m, 2H).

Example 15.N-(4-(4-amino-7-((1r,4r)-4-(hydroxymethyl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-(methoxy-d₃)benzamide(Compound 18)

Step 1. methyl 5-fluoro-2-(methoxy-d₃)benzoate (18b)

To a mixture of methyl 5-fluoro-2-hydroxybenzoate 18a (1.7 g, 10 mmol)and potassium carbonate (2.07 g, 15 mmol) in acetonitrile (10 mL) wasadded deuterated methyl iodide (1.45 g, mmol). After stirring for 24hours, it was filtered. The filtrate was removed from the solvent underreduced pressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=100/1 to 4/1) to give thetitle compound 18b (900 mg, 48%).

MS m/z (ESI): 188 [M+1]

Step 2. 5-fluoro-2-(methoxy-d₃)benzoic acid (18c)

To a solution of 18b (900 mg, 4.81 mmol) in THE (5 mL) was added anaqueous solution of sodium hydroxide (231 mg, 5.77 mmol, 2 mL). Afterstirring for 5 hours, the mixture was adjusted to pH=4 with a solutionof hydrogen chloride in ethyl acetate (4 M) and extracted with ethylacetate (3×10 mL). The combined organic phase was washed with water(3×10 mL), dried over anhydrous sodium sulfate, and filtered. Thefiltrate was removed from the solvent under reduced pressure to give thetitle compound 18c (600 mg, 72%). The product was used directly in thenext step without further purification.

MS m/z (ESI): 173 [M+1]

Step 3. (4-((5-fluoro-2-(methoxy-d₃)benzamido)methyl)phenyl)boronic acid(18d)

To a mixture of 18c (600 mg, 3.47 mmol), (4-(aminomethyl)phenyl)boronicacid hydrochloride (649 mg, 3.47 mmol) and DIPEA (895 mg, 6.93 mmol) inDMF (5 mL) was added HATU (1.58 g, 4.16 mmol). The mixture was stirredfor 18 hours and then purified by prep-HPLC to give the title compound18d (360 mg, 34%).

MS m/z (ESI): 307 [M+1]

Step 4. methyl(1r,4r)-4-(4-amino-5-(4-((5-fluoro-2-(methoxy-d₃)benzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)cyclohexane-1-carboxylate(18e)

A mixture of 18e (50 mg, 0.12 mmol), 18d (46 mg, 0.15 mmol), PdCl₂(dppf)(10 mg, 0.012 mmol), potassium carbonate (26 mg, 0.19 mmol), 1,4-dioxane(2 mL) and water (0.2 mL) was heated to 130° C. under a nitrogenatmosphere and stirred for 1 hour. After cooling to room temperature,the solvent was removed under reduced pressure and the residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=100/1 to 1/100) to give the title compound 18f (50 mg, 75%).

MS m/z (ESI): 536 [M+1]

Step 5.N-(4-(4-amino-7-((1r,4r)-4-(hydroxymethyl)cyclohexyl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-(methoxy-d₃)benzamide(18)

Lithium aluminum hydride (8 mg, 0.19 mmol) was added to a solution of18f (50 mg, 0.093 mmol) in THE (3 mL) at 0° C. The mixture was graduallywarmed to room temperature and stirred for 3 hours. It was quenched withwater (0.1 mL) and filtered. The filtrate was removed from the solventunder reduced pressure and the residue was purified by prep-HPLC to givethe title compound 18 (15.4 mg, solid, 33%).

MS m/z (ESI): 508 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.86 (s, 1H), 7.67-7.61 (m, 3H), 7.58-7.54 (m,2H), 7.30-7.25 (m, 1H), 7.21-7.17 (m, 1H), 4.72 (s, 2H), 3.45 (d, J=6.3Hz, 2H), 2.12-2.06 (m, 2H), 2.02-1.96 (m, 2H), 1.90-1.80 (m, 2H),1.69-1.56 (m, 2H), 1.25-1.15 (m, 2H).

Example 16.N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-3-methoxy-6-methylpicolinamide(Compound 19)

Step 1. (3-methoxy-6-methylpyridin-2-yl)methanol (19b)

To a mixture of 2-(hydroxymethyl)-6-methylpyridin-3-ol 19a (700 mg, 5mmol) and potassium carbonate (2.07 g, 15 mmol) in DMF (10 mL) was addedmethyl iodide (1.42 g, 10 mmol). After stirring for 1 hour, the mixturewas added with water (10 mL) and extracted with ethyl acetate (3×10 mL).The combined organic phase was dried over anhydrous sodium sulfate andthe solvent was removed from the filtrate under reduced pressure to givethe title compound 19b (600 mg, crude). The product was used directly inthe next step without further purification.

MS m/z (ESI): 154 [M+1]

Step 2. 3-methoxy-6-methylpicolinic acid (19c)

To a mixture of 19b (600 mg, 4 mmol) and water (40 mL) were added apotassium hydroxide aqueous solution (1 N, 20 mL) and potassiumpermanganate (1.26 g, 8 mmol) in sequence. After stirring for 1.5 hours,the mixture was adjusted to pH=4-6 with dilute hydrochloric acid (1 N)and extracted with dichloromethane (5×20 mL). The combined organic phasewas concentrated under reduced pressure and the residue was purified bysilica gel column chromatography (dichloromethane/methanol=100/0 to 4/1)to give the title compound 19c (150 mg, 18%).

MS m/z (ESI): 168 [M+1]

Step 3. (4-((3-methoxy-6-methylpicolinamido)methyl)phenyl)boronic acid(19d)

To a mixture of 19c (150 mg, 0.90 mmol), (4-(aminomethyl)phenyl)boronicacid hydrochloride (187 mg, 1.00 mmol) and DIPEA (258 mg, 2.0 mmol) inDMF (2 mL) was added HATU (380 mg, 1.0 mmol). After stirring for onehour, the mixture was added with water (10 mL) and extracted with ethylacetate (3×10 mL). The combined organic phase was dried over anhydroussodium sulfate, filtered, and concentrated under reduced pressure. Theresidue was purified by prep-HPLC to give the title compound 19d (30 mg,11%).

MS m/z (ESI): 301 [M+1]

Step 4.N-(4-(8-((2,4-dimethoxybenzyl)amino)-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-3-methoxy-6-methylpicolinamide(19f)

A mixture of 19e (59 mg, 0.13 mmol), 19d (30 mg, 0.13 mmol), potassiumcarbonate (51 mg, 0.39 mmol), PdCl₂(dppf) (10 mg, 0.013 mmol),1,4-dioxane (3 mL) and water (0.3 mL) was heated to 100° C. under anitrogen atmosphere and stirred for 18 hours. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography (petroleumether/ethyl acetate=100/0 to 3/2) to give the title compound 19f (30 mg,37%).

MS m/z (ESI): 623 [M+1]

Step 5.N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-3-methoxy-6-methylpicolinamide(19)

A mixture of 19f (30 mg, 0.048 mmol) and trifluoroacetic acid (5 mL) washeated to 100° C. and stirred for one hour. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by prep-HPLC to give the title compound 19 (6.0 mg,solid, 27%).

MS m/z (ESI): 473 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.73-7.45 (m, 6H), 7.39 (s, 1H), 7.00 (d,J=4.9 Hz, 1H), 4.70 (s, 2H), 4.11-3.85 (m, 5H), 3.71 (t, J=10.9 Hz, 1H),3.53 (t, J=10.6 Hz, 1H), 3.44 (t, J=11.1 Hz, 1H), 2.48 (s, 3H), 2.15 (d,J=11.9 Hz, 1H), 2.09-1.93 (m, 1H), 1.89 (dd, J=23.3, 10.2 Hz, 1H), 1.77(d, J=13.1 Hz, 1H).

Example 17.N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-methoxy-2-methylisonicotinamide(Compound 20)

Step 1. 2-bromo-5-methoxyisonicotinic acid (20b)

To a solution of 2-bromo-5-fluoroisonicotinic acid 20a (2.2 g, 10 mmol)in methanol (40 mL) was added sodium methoxide (2.7 g, 30% in methanol,15 mmol). The mixture was heated to 60° C. and stirred for 4 hours.After cooling to room temperature, sodium methoxide (1.8 g, 30% inmethanol, 10 mmol) was added and the mixture was heated to 60° C. againand stirred for 18 hours. After cooling to room temperature, the solventwas removed under reduced pressure. The residue was dissolved in water(20 mL) and the mixture was adjusted to pH=4 with hydrochloric acid (12N). After filtration, the precipitate was collected to give the titlecompound 20b (2.06 g, 89%).

MS m/z (ESI): 232 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 13.62 (s, 1H), 8.34 (s, 1H), 7.70 (s, 1H),3.92 (s, 3H).

Step 2. methyl 2-bromo-5-methoxyisonicotinate (20c)

To a solution of 20b (2.06 g, 8.9 mmol) in methanol (60 mL) was addedconcentrated sulfuric acid (1.13 g, 1.3 mmol) and the mixture was heatedto reflux for 16 hours. After cooling to room temperature, the solventwas removed under reduced pressure, and to which was added saturatedsodium bicarbonate solution (100 mL) The mixture was then extracted withdichloromethane (3×50 mL). The combined organic phase was dried overanhydrous sodium sulfate, filtered, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(petroleum ether/ethyl acetate=100/0 to 2/3) to give the title compound20c (1.86 g, 85%).

MS m/z (ESI): 246 [M+1]

¹H NMR (400 MHz, CDCl₃) δ 8.19 (s, 1H), 7.74 (s, 1H), 3.99 (s, 3H), 3.93(s, 3H).

Step 3. methyl 5-methoxy-2-methylisonicotinate (20d)

To solution of 20c (1.61 g, 6.6 mmol) in THE (24 mL) were addedtrimethylaluminum (2 M in hexane, 3.94 mL, 7.9 mmol) and palladiumtetrakis(triphenylphosphine) (758 mg, 0.66 mmol) under nitrogenatmosphere. The mixture was heated to 70° C. and stirred for 4 hours.After cooling to room temperature, saturated ammonium chloride solution(100 mL) and ethyl acetate (100 mL) were added sequentially. Theprecipitate was filtered out and the filtrate was allowed to stand. Theseparated aqueous layer was extracted with ethyl acetate (2×100 mL). Thecombined organic phase was washed with saturated brine (100 mL), driedover anhydrous sodium sulfate, filtered, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(dichloromethane/ethyl acetate=100/0 to 2/3) to give the title compound20d (1.43 g, mixed with triphenylphosphine). The product was useddirectly in the next step without further purification.

MS m/z (ESI): 182 [M+1]

¹H NMR (400 MHz, CDCl₃) δ 8.32 (s, 1H), 7.44 (s, 1H), 3.98 (s, 3H), 3.92(s, 3H), 2.54 (s, 3H).

Step 4. 5-methoxy-2-methylisonicotinic acid (20e)

To a mixture of 20d (1.43 g, mixed with triphenylphosphine) in methanol(6 mL) and THE (6 mL) was added sodium hydroxide solution (2.5 N, 6 mL,15 mmol). The mixture was stirred for 16 hours, and the solvent wasremoved under reduced pressure. The residue was dissolved in water (20mL) and washed with isopropyl ether (3×20 mL). The aqueous layer wasacidified with concentrated hydrochloric acid to pH=4, concentrated to avolume of about 2 mL and extracted with THF (100 mL). The organic phasewas dried over anhydrous sodium sulfate, filtered, and concentratedunder reduced pressure to give the title compound 20e (633 mg, 50% overtwo steps).

MS m/z (ESI): 168 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 13.27 (s, 1H), 8.36 (s, 1H), 7.36 (s, 1H),3.88 (s, 3H), 2.43 (s, 3H).

Step 5. (4-((5-methoxy-2-methylisonicotinamido)methyl)phenyl)boronicacid (20f)

To a mixture of 20e (119 mg, 0.71 mmol) and(4-(aminomethyl)phenyl)boronic acid hydrochloride (133 mg, 0.71 mmol) indichloromethane (2 mL) were added DIPEA (367 mg, 2.84 mmol) and HATU(270 mg, 0.71 mmol). The mixture was stirred for one hour and thesolvent was removed under reduced pressure. The residue was purified bysilica gel column chromatography (dichloromethane/methanol=100/0 to19/1) to give the title compound 20f (80 mg, 38%).

MS m/z (ESI): 301 [M+1]

Step 6.N-(4-(8-((2,4-dimethoxybenzyl)amino)-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-methoxy-2-methylisonicotinamide(20g)

A mixture of 20f (80 mg, 0.27 mmol), 19e (119 mg, 0.27 mmol), potassiumcarbonate (149 mg, 1.1 mmol), PdCl₂(dppf) (20 mg, 0.027 mmol),1,4-dioxane (20 mL) and water (2 mL) was heated to 100° C. and stirredfor 8 hours under a nitrogen atmosphere. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography(dichloromethane/methanol=100/0 to 26/1) to give the title compound 20g(54 mg, 32%).

MS m/z (ESI): 623 [M+1]

Step 7.N-(4-(8-amino-3-(tetrahydro-2H-pyran-3-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-methoxy-2-methylisonicotinamide(20)

A solution of 20g (54 mg, 0.087 mmol) in trifluoroacetic acid (5 mL) washeated to 80° C. and stirred for 20 hours. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by prep-HPLC to give the title compound 20 (10.8mg, solid, 26%).

MS m/z (ESI): 473 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 8.36 (s, 1H), 7.70-7.52 (m, 6H), 7.04 (d,J=4.7 Hz, 1H), 4.72 (s, 2H), 4.13-3.99 (m, 2H), 4.07 (s, 3H), 3.74 (t,J=10.9 Hz, 1H), 3.56 (td, J=11.5, 2.5 Hz, 1H), 3.51-3.43 (m, 1H), 2.54(s, 3H), 2.18 (d, J=12.7 Hz, 1H), 2.04 (ddd, J=15.7, 12.4, 4.2 Hz, 1H),1.95-1.84 (m, 1H), 1.83-1.75 (m, 1H).

Compounds as shown below were synthesized according to the proceduresfor the sixth step to the seventh step in Example 20, except that adifferent phenylboronic acid was used instead of(4-((5-methoxy-2-methylisonicotinamido)methyl)phenyl)boronic acid 20f inthe sixth step.

Compound MS m/z No. Structure Replacement of 20f (ESI) 21

1b 476 25

18d 479

The NMR data of compounds 21 and 25 are shown below:

Compound ¹H NMR N-(4-(8-amino-3- ¹H NMR (400 MHz, DMSO-d₆) δ12.68 (s,1H), 8.85 (t, J = 6.0 (tetrahydro-2H-pyran- Hz, 1H), 8.13 (s, 1H) 7.70(d, J = 5.1 Hz, 1H), 7.58-7.49 (m, 3-yl)imidazo[1,5- 3H), 7.45 (d, J =8.1 Hz, 2H), 7.39-7.30 (m, 1H), 7.19 (dd, J = a]pyrazin-1-yl)benzyl)-9.1, 4.3 Hz, 1H), 7.03 (d, J = 5.0 Hz, 1H), 5.99 (s, 2H), 4.575-fluoro-2- (d, J = 6.1 Hz, 2H), 4.00 (d, J = 11.0 Hz, 1H), 3.90 (s,3H), methoxybenzamide (21) 3.58 (t, J = 10.8 Hz, 1H), 3.47-3.36 (m, 2H),2.08 (d, J = 11.2 Hz, 1H), 1.94-1.65 (m, 4H). N-(4-(8-amino-3- ¹H NMR(400 MHz, DMSO-d₆) δ 8.85 (t, J = 5.9 Hz, 1H), (tetrahydro-2H-pyran-7.70 (d, J = 5.0 Hz, 1H), 7.60-7.40 (m, 5H), 7.37-7.31 (m,3-yl)imidazo[1,5- 1H), 7.18 (dd, J = 9.1, 4.2 Hz, 1H), 7.02 (d, J = 4.9Hz, 1H), a]pyrazin-1-yl)benzyl)- 5.98 (s, 2H), 4.57 (d, J = 6.0 Hz, 2H),3.95 (dd, J = 35.6, 10.9 5-fluoro-2-(methoxy- Hz, 2H), 3.59 (t, J = 10.8Hz, 1H), 3.46-3.36 (m, 2H), 2.08 d₃)benzamide (25) (d, J = 7.8 Hz, 1H),1.92-1.69 (m, 3H).

Example 18.(R)—N-(4-(4-amino-7-(pyrrolidin-3-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 22)

Step 1. 1-benzyl 3-(2,5-dioxopyrrolidin-1-yl)(R)-pyrrolidine-1,3-dicarboxylate (22b)

A mixture of (R)-1-((benzyloxy)carbonyl)pyrrolidine-3-carboxylic acid22a (748 mg, 3 mmol) and 1-hydroxy-pyrrolidine-2,5-dione (380 mg, 3.3mmol) in THE (30 mL) was cooled to 0° C. and added withdicyclohexylcarbodiimide (743 mg, 3.6 mmol). After warming to roomtemperature and stirring for 4 hours, the mixture was cooled to 0° C.and filtered. The filtrate was concentrated to dryness under reducedpressure to give the title compound 22b (1.1 g, 100%). The product wasused directly in the next step without further purification.

MS m/z (ESI): 347 [M+1]

Step 2. benzyl(R)-3-(((5-oxo-4,5-dihydro-1,2,4-triazin-6-yl)methyl)carbamoyl)pyrrolidine-1-carboxylate(22c)

To a mixture of 12f (915 mg, 6 mmol) and DBU (4.6 g, 30 mmol) in DMF (10mL) was added 22b (1.1 g, 3 mmol) in THE (10 mL). After stirring for 4hours, the solvent was removed under reduced pressure and the residuewas purified by prep-HPLC to give the title compound 22c (240 mg, 22%).

MS m/z (ESI): 358 [M+1]

Step 3. benzyl(R)-3-(4-oxo-3,4-dihydroimidazo[5,1-f][1,2,4]triazin-7-yl)pyrrolidine-1-carboxylate(22d)

To a solution of 22c (240 mg, 0.67 mmol) in acetonitrile (15 mL) wasadded phosphine oxychloride (5 mL). The resulting mixture was heated to80° C. and stirred for 5 hours. After cooling to room temperature, thesolvent was removed under reduced pressure to give the title compound22d (270 mg, crude). The product was used directly in the next stepwithout further purification.

MS m/z (ESI): 340 [M+1]

Step 4. benzyl(R)-3-(5-iodo-4-oxo-3,4-dihydroimidazo[5,1-f][1,2,4]triazin-7-yl)pyrrolidine-1-carboxylate(22e)

To a solution of 22d (270 mg, crude, 0.67 mmol) in DMF (10 mL) was addedNIS (1.51 g, 6.7 mmol). The resulting mixture was heated to 50° C. andstirred for 2 hours. After cooling to room temperature, the solvent wasremoved under reduced pressure and the residue was purified by prep-HPLCto give the title compound 22e (140 mg, 45%).

MS m/z (ESI): 466 [M+1]

Step 5. benzyl(R)-3-(4-amino-5-iodoimidazo[5,1-J][1,2,4]triazin-7-yl)pyrrolidine-1-carboxylate(22f)

To a solution of 1H-[1,2,4]triazole (207 mg, 3 mmol) in pyridine (1 mL)was added phosphine oxychloride (138 mg, 0.9 mmol). After stirring for10 minutes, a solution of 22e (140 mg, 0.3 mmol) in pyridine (1 mL) wasadded. After stirring for additional 2 hours, a solution of ammonia inisopropanol (2 M, 5 mL, 10 mmol) was added and stirring was continuedfor another hour. The solvent was removed under reduced pressure and theresidue was purified by silica gel column chromatography (petroleumether/ethyl acetate=20/1 to 1/2) to give the title compound 22f (220 mg,crude). The product was used directly in the next step without furtherpurification.

MS m/z (ESI): 465 [M+1]

Step 6. benzyl(R)-3-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)pyrrolidine-1-carboxylate(22g)

A mixture of 22f (220 mg, crude product), 1b (136 mg, 0.45 mmol),potassium carbonate (83 mg, 0.6 mmol), PdCl₂(dppf) (44 mg, 0.06 mmol),1,4-dioxane (4 mL) and water (1 mL) were heated to 105° C. under anitrogen atmosphere and stirred for 1.5 hours. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by prep-HPLC to give the title compound 22g (45 mg,25%).

MS m/z (ESI): 596 [M+1]

Step 7.(R)—N-(4-(4-amino-7-(pyrrolidin-3-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(22)

To a solution of 22g (45 mg, 0.076 mmol) in dichloromethane (10 mL) at0° C. was added trimethylsilyl iodide (30 mg, 0.15 mmol). The mixturewas warmed to room temperature and stirred for 2 hours. The solvent wasremoved under reduced pressure and the residue was purified by prep-HPLCto give the title compound 22 (20 mg, solid, 57%).

MS m/z (ESI): 462 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.90 (s, 1H), 7.67-7.59 (m, 3H), 7.55 (d,J=8.3 Hz, 2H), 7.26 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.18 (dd, J=9.1, 4.2Hz, 1H), 4.69 (s, 2H), 4.32-4.25 (m, 1H), 3.97 (d, J=8.2 Hz, 3H),3.81-3.73 (m, 2H), 3.60-3.54 (m, 1H), 3.47 (dd, J=7.4, 4.2 Hz, 1H), 2.57(dd, J=13.4, 6.3 Hz, 1H), 2.46-2.39 (m, 1H).

Compounds shown below were synthesized according to the procedures forthe first to sixth steps in Example 18, except that a differentcarboxylic acid was used instead of(R)-1-((benzyloxy)carbonyl)pyrrolidine-3-carboxylic acid 22a in thefirst step.

Compound replacing MS m/z Compound No. Structure 22a (ESI) 26

489 28

559 29

517 43

501

The NMR data of compounds 26, 28, 29 and 43 are shown below:

Compound ¹H NMR N-(4-(4-amino-7-(1,1,1- ¹H NMR (400 MHz, CD₃OD) δ 7.91(s, 1H), 7.65- trifluoropropan-2-yl)imidazo[5,1- 7.61 (m, 3H), 7.55 (d,J = 8.3 Hz, 2H), 7.26 f][1,2,4]triazin-5-yl)benzyl)-5- (ddd, J = 9.1,7.6, 3.3 Hz, 1H), 7.17 (dd, J = 9.1, fluoro-2-methoxybenzamide (26) 4.2Hz, 1H), 4.70 (s, 2H), 4.58-4.51 (m, 1H), 3.97 (s, 3H), 1.66 (d, J = 7.3Hz, 3H). methyl 4-(4-amino-5-(4-((5-fluoro-2- ¹H NMR (400 MHz, CD₃OD) δ7.80 (s, 1H), 7.64- methoxybenzamido)methyl) 7.57 (m, 3H), 7.52 (d, J =8.3 Hz, 2H), 7.25 phenyl)imidazo[5,1-f][1,2,4]triazin- (ddd, J = 9.1,7.6, 3.3 Hz, 1H), 7.17 (dd, J = 9.1, 7-yl)bicyclo[2.2.2]octane-1- 4.2Hz, 1H), 4.69 (s, 2H), 3.96 (s, 3H), 3.66 (s, carboxylate (28) 3H), 2.26(dd, J = 9.7, 6.2 Hz, 6H), 1.93 (dd, J = 9.7, 6.3 Hz, 6H). methyl3-(4-amino-5-(4-((5-fluoro-2- ¹H NMR (400 MHz, CD₃OD) δ7.85 (s, 1H),7.65- methoxybenzamido)methyl)phenyl) 7.59 (m, 3H), 7.54 (d, J = 8.3 Hz,2H), 7.28- imidazo[5,1-f][1,2,4]triazin-7- 7.22 (m, 1H), 7.17 (dd, J =9.1, 4.2 Hz, 1H), 4.69 yl)bicyclo[1.1.1]pentane-1- (s, 2H), 3.97 (s,3H), 3.71 (s, 3H), 2.63 (s, 6H). carboxylate (29) N-(4-(4-amino-7-(1- ¹HNMR (400 MHz, CD₃OD) δ 7.93 (s, 1H), 7.62 (trifluoromethyl) (dd, J =8.7, 2.4 Hz, 3H), 7.55 (d, J = 8.1 Hz, cyclopropyl)imidazo[5,1- 2H),7.28-7.22 (m, 1H), 7.17 (dd, J = 9.1, 4.2f][1,2,4]triazin-5-yl)benzyl)-5- Hz, 1H), 4.70 (s, 2H), 3.97 (s, 3H),1.59 (dd, J = fluoro-2-methoxybenzamide (43) 7.6, 5.5 Hz, 2H), 1.43 (d,J = 6.0 Hz, 2H).

Example 19.N-(4-(8-amino-3-((1r,4r)-4-(methylcarbamoyl)cyclohexyl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 23)

To a mixture of 13 (8 mg, 0.015 mmol), ammonium chloride (3 mg, 0.045mmol) and DIPEA (8 mg, 0.06 mmol) in DMF (2 mL) was added HATU (11 mg,0.03 mmol). After stirring for 30 minutes, the solvent was removed underreduced pressure and the residue was purified by prep-HPLC to give thetitle compound 23 as a formate salt (5.5 mg, solid, 63%).

MS m/z (ESI): 531 [M+1]

1H NMR (400 MHz, CD₃OD) δ 9.02 (s, 1H), 7.88 (d, J=4.9 Hz, 1H), 7.71 (d,J=5.6 Hz, 1H), 7.60 (dt, J=24.3, 6.5 Hz, 4H), 7.29-7.22 (m, 1H), 7.18(dd, J=9.1, 4.2 Hz, 1H), 6.98 (d, J=5.5 Hz, 1H), 4.71 (d, J=3.9 Hz, 2H),3.98 (s, 3H), 3.22-3.14 (m, 1H), 2.73 (d, J=4.5 Hz, 3H), 2.31 (dd,J=13.4, 9.8 Hz, 1H), 2.08 (d, J=13.4 Hz, 2H), 1.96 (d, J=12.4 Hz, 2H),1.87-1.65 (m, 4H).

Example 20.(R)—N-(4-(4-amino-7-(1-(cyclopropanecarbonyl)pyrrolidin-3-yl)imidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 24)

To a mixture of 22 (110 mg, 0.24 mmol) and saturated sodium bicarbonatesolution (5 mL) in THE (10 mL) at 0° C. was added cyclopropylformylchloride (37 mg, 0.36 mmol). After warming to room temperature, themixture was stirred for 30 minutes, diluted with saturated brine (10 mL)and extracted with ethyl acetate (2×50 mL). The combined organic phasewas dried over anhydrous sodium sulfate, filtered, and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (dichloromethane/methanol=100/1 to 15/1) to give thetitle compound 24 (57.5 mg, solid, 45%).

MS m/z (ESI): 530 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.87 (dd, J=6.5, 2.7 Hz, 1H), 7.62 (dd, J=9.1,3.0 Hz, 3H), 7.53 (d, J=8.1 Hz, 2H), 7.28-7.22 (m, 1H), 7.17 (dd, J=9.1,4.2 Hz, 1H), 4.69 (s, 2H), 4.30-4.19 (m, 1H), 4.11-3.99 (m, 2H), 3.97(s, 3H), 3.91-3.68 (m, 2H), 2.55-2.37 (m, 2H), 1.87-1.78 (m, 1H),0.93-0.81 (m, 4H).

Example 21.1-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-4-methylpiperidine-4-carboxylicacid (Compound 27)

Step 1. methyl1-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-4-methylpiperidine-4-carboxylate(27b)

A mixture of 27a (360 mg, 0.52 mmol) and trifluoroacetic acid (5 mL) washeated to 90° C. and stirred for 5 hours. After cooling to roomtemperature, the solvent was removed under reduced pressure and theresidue was purified by prep-HPLC to give the title compound 27b (180mg, 64%).

MS m/z (ESI): 547 [M+1]

Step 2.1-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-4-methylpiperidine-4-carboxylicacid (27)

A mixture of 27b (180 mg, 0.33 mmol), water (2 mL), methanol (2 mL) andlithium hydroxide (79 mg, 3.3 mmol) was stirred for two hours. Thesolvent was then removed under reduced pressure and the residue waspurified by prep-HPLC to give the title compound 27 (21.2 mg, solid,12%).

MS m/z (ESI): 533 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 12.40 (s, 1H), 8.84 (t, J=5.6 Hz, 1H), 7.57(d, J=8.2 Hz, 2H), 7.52 (dd, J=9.2, 3.3 Hz, 1H), 7.44 (d, J=8.2 Hz, 2H),7.37-7.31 (m, 1H), 7.23 (d, J=5.0 Hz, 1H), 7.19 (dd, J=9.1, 4.3 Hz, 1H),6.95 (d, J=5.0 Hz, 1H), 6.01 (s, 2H), 4.56 (d, J=6.0 Hz, 2H), 3.90 (s,3H), 3.22 (m, 2H), 2.96 (m, 2H), 2.11 (d, J=13.4 Hz, 2H), 1.63 (m, 2H),1.22 (s, 3H).

Example 22.3-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)bicyclo[1.1.1]pentane-1-carboxylicacid (Compound 31)

A mixture of 29 (65 mg, 0.13 mmol) and aqueous lithium hydroxide (1 N, 2mL) in THF (10 mL) was stirred for four hours. The mixture was adjustedto pH=7-8 with glacial acetic acid and extracted with ethyl acetate(2×50 mL). The combined organic phase was dried over anhydrous sodiumsulfate, filtered, and concentrated under reduced pressure. The residuewas purified by prep-HPLC to give the title compound 31 (50 mg, solid,79%).

MS m/z (ESI): 503 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.86 (s, 1H), 7.64-7.59 (m, 3H), 7.54 (d,J=8.3 Hz, 2H), 7.25 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.17 (dd, J=9.1, 4.2Hz, 1H), 4.70 (d, J=4.0 Hz, 2H), 3.97 (s, 3H), 2.62 (s, 6H).

Example 23.(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-1,4-dimethylcyclohexane-1-carboxylicacid (Compound 32)

Step 1. dimethyl 1,4-dimethylcyclohexane-1,4-dicarboxylate (32b)

To a solution of LDA (2 M, 12.5 mL, 25 mmol) and hexamethylphosphorictriamide (13.9 mL, 79.9 mmol) in THE (25 mL) at −78° C. was addeddimethyl cyclohexane-1,4-dicarboxylate 32a (2 g, 10 mmol) in THE (2 mL).After stirring for one hour, the mixture was warmed to 0° C. and stirredfor another hour. The mixture was re-cooled to −78° C. and methyl iodide(7.75 g, 54.6 mmol) was added. The resulting mixture was stirred at roomtemperature for two hours, added with 1 N hydrochloric acid (3 mL) andextracted with ethyl acetate (50 mL). The combined organic phase wasdried over anhydrous sodium sulfate, filtered, and concentrated todryness under reduced pressure. The residue was purified by silica gelcolumn chromatography (petroleum ether/ethyl acetate=78/22) to give thetitle compound 32b (450 mg, 20%).

¹H NMR (400 MHz, DMSO-d₆) δ 3.59 (s, 6H), 1.79-1.70 (m, 4H), 1.47-1.38(m, 4H), 1.13 (s, 6H).

Step 2. 4-(methoxycarbonyl)-1,4-dimethylcyclohexane-1-carboxylic acid(32c)

A mixture of 32b (387 mg, 1.70 mmol), potassium hydroxide (476 mg, 8.5mmol), methanol (2 mL) and water (2 mL) were heated to 80° C. andstirred for 16 hours. After cooling to room temperature, the reactionmixture was concentrated to dryness under reduced pressure, added with 1N hydrochloric acid (5 mL) and extracted with ethyl acetate (8 mL). Thecombined organic phase was dried over anhydrous sodium sulfate,filtered, and concentrated to dryness under reduced pressure to give thetitle compound 32c (325 mg, crude, 90%).

¹H NMR (400 MHz, DMSO-d₆) δ 3.59 (s, 3H), 1.82-1.70 (m, 4H), 1.46-1.32(m, 4H), 1.12 (d, J=7.6 Hz, 6H).

Step 3. methyl(1r,4r)-4-(((3-chloropyrazin-2-yl)methyl)carbamoyl)-1,4-dimethylcyclohexane-1-carboxylate(32d) and methyl(1s,4s)-4-(((3-chloropyrazin-2-yl)methyl)carbamoyl)-1,4-dimethylcyclohexane-1-carboxylate(32e)

To a mixture of 32c (492 mg, 2.30 mmol) and(3-chloro-pyrazin-2-yl)methylamine hydrochloride (621 mg, 3.45 mmol) inDMF (15 mL) were added DIPEA (1.48 g, 11.45 mmol) and HATU (1.31 g, 15mL). After stirring for 2 hours, the mixture was purified by prep-HPLCand then by silica gel column chromatography (petroleum ether/ethylacetate=1/1) to give the title compound 32d (190 mg, 24%) and 32e (60mg, 8%)

MS m/z (ESI): 340 [M+1]

Step 4 through Step 7. methyl(1r,4r)-4-(8-((2,4-dimethoxybenzyl)amino)-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-1,4-dimethylcyclohexane-1-carboxylate(32j)

32j was synthesized according to the procedures for step 2 to 5 inExample 4, except that 32d was used instead of 5b.

MS m/z (ESI): 710 [M+1]

Step 8. methyl(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-1,4-dimethylcyclohexane-1-carboxylate(32k)

A solution of 32j (60 mg, 0.084 mmol) in trifluoroacetic acid (3 mL) washeated to 90° C. and stirred for 5 hours. After cooling to roomtemperature, the mixture was concentrated to dryness and the residue waspurified by prep-HPLC to give the title compound 32k (30 mg, 63%).

MS m/z (ESI): 560 [M+1]

Step 9.(1r,4r)-4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-1,4-dimethylcyclohexane-1-carboxylicacid (32)

A mixture of 32k (30 mg, 0.054 mmol), lithium hydroxide monohydrate (9.6mg, 0.24 mmol), water (1 mL) and methanol (1 mL) was stirred for 2 hoursand then concentrated to dryness under reduced pressure. The residue waspurified by prep-HPLC to give the title compound 32 (9 mg, solid, 31%).

MS m/z (ESI): 546 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 8.24 (s, 1H), 7.73 (d, J=5.5 Hz, 1H), 7.63(dd, J=9.0, 3.1 Hz, 3H), 7.54 (d, J=8.3 Hz, 2H), 7.26 (ddd, J=9.1, 7.6,3.3 Hz, 1H), 7.17 (dd, J=9.1, 4.2 Hz, 1H), 6.94 (d, J=5.4 Hz, 1H), 4.70(s, 2H), 3.97 (s, 3H), 2.58 (d, J=14.4 Hz, 2H), 2.09 (d, J=13.8 Hz, 2H),1.65 (t, J=11.9 Hz, 2H), 1.39 (s, 5H), 1.09 (s, 3H).

Compound 39 was synthesized according to the procedures for the fourthto eighth step in Example 23, except that 32e was used instead of 32d inthe fourth step.

Compound replacing MS m/z Compound No. Structure 32d (ESI) 39

32e 546

The NMR data of Compound 39 is shown below:

Compound ¹H NMR methyl (1s,4s)-4-(8-amino-1- ¹H NMR (400 MHz, DMSO-d₆) δ8.86 (4-((5-fluoro-2- (s, 1H), 7.79 (d, J = 5.5 Hz, 1H), 7.53methoxybenzamido)methyl) (m, 5H), 7.38-7.30 (m, 1H), 7.19 (m, 1H),phenyl)imidazo[1,5-a]pyrazin- 6.98 (d, J = 5.5 Hz, 1H), 6.85 (s, 2H),3-yl)-1,4- 4.58 (d, J = 6.1 Hz, 2H), 3.90 (s, 3H), dimethylcyclohexane-3.57 (s, 3H), 2.08 (t, J = 10.4 Hz, 2H), 1-carboxylate (39) 2.01-1.81(m, 4H), 1.53 (s, 2H), 1.43 (s, 3H), 1.21 (s, 3H).

Example 24. methyl4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylate(Compound 33)

Step 1. methyl4-(((3-chloropyrazin-2-yl)methyl)carbamoyl)bicyclo[2.2.2]octane-1-carboxylate(33b)

To a mixture of 4-(methoxycarbonyl)bicyclo[2.2.2]octane-1-carboxylicacid 33a (1.06 g, 5 mmol), (3-chloropyrazin-2-yl)methylaminehydrochloride 5a (900 mg, 5 mmol) and DIPEA (1.94 g, 15 mmol) in DMF (10mL) was added HATU (2.47 g, 6.5 mmol). After stirring for 30 minutes,the mixture was diluted with water (40 mL) and extracted with ethylacetate (2×100 mL). The combined organic phase was washed with saturatedbrine (80 mL), dried over anhydrous sodium sulfate, filtered, andconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (petroleum ether/ethyl acetate=50/1 to 2/1) togive the title compound 33b (1.5 g, 89%).

MS m/z (ESI): 338 [M+1]

Step 2. methyl4-(8-chloroimidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylate(33c)

A mixture of 33b (1.5 g, 4.44 mmol) and phosphine oxychloride (20 mL) inacetonitrile (10 mL) was heated to 120° C. and stirred for 4 hours.After cooling to room temperature, the solvent was removed under reducedpressure to give the title compound 33c (1.4 g, crude). The product wasused directly in the next step without further purification.

MS m/z (ESI): 320 [M+1]

Step 3. methyl4-(8-chloro-1-iodoimidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylate(33d)

To a mixture of 33c (1.4 g, crude product) in DMF (10 mL) was added NIS(2 g, 8.9 mmol). The resulting mixture was heated to 50° C. and stirredfor 2 hours. After cooling to room temperature, the mixture was dilutedwith water (30 mL) and extracted with ethyl acetate (2×100 mL). Thecombined organic phase was washed with saturated brine (80 mL), driedover anhydrous sodium sulfate, filtered, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(petroleum ether/ethyl acetate=50/1 to 2/1) to give the title compound33d (1.5 g, 76% in two steps).

MS m/z (ESI): 446 [M+1]

Step 4. methyl4-(8-((2,4-dimethoxybenzyl)amino)-1-iodoimidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylate(33e)

A mixture of 33d (1.5 g, 3.4 mmol), 2,4-dimethoxybenzylamine (675 mg,4.0 mmol) and DIPEA (868 mg, 6.7 mmol) in acetonitrile (30 mL) washeated at 90° C. and stirred for 15 hours in a sealed tube. Aftercooling to room temperature, the solvent was removed under reducedpressure and the residue was purified by silica gel columnchromatography (petroleum ether/ethyl acetate=50/1 to 3/1) to give thetitle compound 33e (1.7 g, 88%).

MS m/z (ESI): 577 [M+1]

Step 5. methyl4-(8-((2,4-dimethoxybenzyl)amino)-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylate(33f)

A mixture of 33e (1.7 g, 3 mmol), 1b (1.36 g, 4.5 mmol), potassiumcarbonate (828 mg, 6 mmol), PdCl₂(dppf) (219 mg, 0.3 mmol), 1,4-dioxane(16 mL) and water (4 mL) was heated to 100° C. under a nitrogenatmosphere and stirred for 2 hours. After cooling to room temperature,the solvent was removed under reduced pressure and the residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=50/1 to 1/2) to give the title compound 33f (1.7 g, 80%).

MS m/z (ESI): 708 [M+1]

Step 6. methyl4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylate(33)

A mixture of 33f (1.7 g, 2.4 mmol), dichloromethane (5 mL) andtrifluoroacetic acid (30 mL) was heated to reflux and stirred for 5hours. After cooling to room temperature, the solvent was removed underreduced pressure, and the residue was dissolved in methanol (50 mL) andadjusted to pH 7-8 with saturated sodium bicarbonate solution. Thesolvent was then removed under reduced pressure and the residue waspurified by silica gel column chromatography(dichloromethane/methanol=100/1 to 20/1) to give the title compound 33(1.2 g, solid, 89%).

MS m/z (ESI): 558 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.80 (d, J=5.3 Hz, 1H), 7.62 (dd, J=9.2, 3.2Hz, 1H), 7.55 (dd, J=19.7, 8.3 Hz, 4H), 7.28-7.22 (m, 1H), 7.17 (dd,J=9.1, 4.2 Hz, 1H), 6.93 (d, J=5.3 Hz, 1H), 4.69 (s, 2H), 3.96 (s, 3H),3.67 (s, 3H), 2.18 (dd, J=9.8, 5.9 Hz, 6H), 1.98 (dd, J=9.7, 5.9 Hz,6H).

The compounds as shown below were synthesized according to theprocedures in Example 24, except that a different carboxylic acid wasused instead of 4-(methoxycarbonyl)bicyclo[2.2.2]octane-1-carboxylicacid 33a in the first step.

Compound MS m/z No. Structure Compound replacing 33a (ESI) 38

544 42

500

The NMR data of compounds 38 and 42 are shown below:

Compound ¹H NMR methyl 4-(8-amino-1-(4-((5- ¹H NMR (400 MHz, DMSO-d₆) δ8.85 (s, 1H), 7.68 fluoro-2-methoxybenzamido) (d, J = 5.1 Hz, 1H),7.58-7.49 (m, 3H), 7.45 (d, J = methyl)phenyl)imidazo[1,5- 8.2 Hz, 2H),7.37-7.30 (m, 1H), 7.19 (m, 1H), 6.99 a]pyrazin-3- (d, J = 5.0 Hz, 1H),5.96 (s, 2H), 4.57 (d, J = 6.1 Hz, yl)bicyclo[2.2.1]heptane-1- 2H), 3.90(s, 3H), 3.65 (s, 3H), 2.19 (s, 2H), 2.09 (m, carboxylate (38) 6H), 1.77(m, 2H). N-(4-(8-amino-3-(1- ¹H NMR (400 MHz, CD₃OD) δ 7.69 (d, J = 5.1Hz, (trifluoromethyl)cyclopropyl) 1H), 7.66-7.58 (m, 3H), 7.54 (d, J =8.2 Hz, 2H), imidazo[1,5-a]pyrazin-1- 7.25 (ddd, J = 9.1, 7.7, 3.3 Hz,1H), 7.20-7.11 (m, yl)benzyl)-5-fluoro-2- 2H), 4.70 (s, 2H), 3.97 (s,3H), 1.66 (dd, J = 7.3, 5.4 methoxybenzamide (42) Hz, 2H), 1.41 (s, 2H).

Example 25.4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)bicyclo[2.2.2]octane-1-carboxylicacid (Compound 34)

A mixture of 33 (1 g, 1.8 mmol) and aqueous lithium hydroxide solution(1 N, 6 mL) in THE (20 mL) was heated to 50° C. and stirred for 15hours. After cooling to room temperature, glacial acetic acid (1 mL) wasadded to the mixture and the solvent was removed under reduced pressure.The residue was purified by prep-HPLC to give the title compound 34 (300mg, solid, 31%).

MS m/z (ESI): 544 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.81 (d, J=5.3 Hz, 1H), 7.62 (dd, J=9.2, 3.2Hz, 1H), 7.55 (dd, J=20.5, 8.3 Hz, 4H), 7.28-7.23 (m, 1H), 7.17 (dd,J=9.1, 4.2 Hz, 1H), 6.93 (d, J=5.3 Hz, 1H), 4.70 (s, 2H), 3.96 (s, 3H),2.18 (dd, J=9.6, 5.9 Hz, 6H), 1.99 (dd, J=9.8, 5.8 Hz, 6H).

The compounds as shown below were synthesized according to theprocedures in Example 25, except that different esters were used insteadof 33.

Compound replacing MS m/z Compound No. Structure 33 (ESI) 40

38 530 41

39 546 48

48a 561

The NMR data of compounds 40, 41 and 48 are shown below:

Compound ¹H NMR 4-(8-amino-1-(4-((5-fluoro-2- ¹H NMR (400 MHz, DMSO-d₆)δ 12.27 (s, 1H), 8.85 methoxybenzamido)meth- (t, J = 6.1 Hz, 1H), 7.68(d, J = 4.6 Hz, 1H), 7.58- yl)phenyl)imidazo[1,5-a]pyrazin-3- 7.50 (m,3H), 7.45 (d, J = 8.2 Hz, 2H), 7.34 (m, 1H), yl)bicyclo[2.2.1]heptane-1-7.19 (m, 1H), 6.99 (s, 1H), 5.96 (s, 2H), 4.57 (d, J = carboxylic acid(40) 6.1 Hz, 2H), 3.90 (s, 3H), 2.09 (m, 8H), 1.73 (d, J = 9.2 Hz, 2H).(1s,4s)-4-(8-amino-1-(4-((5-fluoro- ¹H NMR (400 MHz, DMSO-d₆) δ 12.16(s, 1H), 8.84 2-methoxybenzamido)meth- (s, 1H), 7.69 (d, J = 5.2 Hz,1H), 7.58-7.50 (m, 3H), yl)phenyl)imidazo[1,5-a]pyrazin-3-yl)- 7.45 (d,J = 8.1 Hz, 2H), 7.33 (m, 1H), 7.19 (m, 1H), 1,4-dimethylcyclohexane-1-6.95 (d, J = 5.1 Hz, 1H), 5.95 (s, 2H), 4.57 (d, J = 6.1 carboxylic acid(41) Hz, 2H), 3.90 (s, 3H), 2.14 (s, 2H), 1.99-1.88 (m, 2H), 1.81 (d, J= 13.8 Hz, 2H), 1.53-1.37 (m, 5H), 1.19 (s, 3H).4-(6-amino-7-(4-((5-fluoro-2- ¹H NMR (400 MHz, CD₃OD) δ 8.10 (s, 1H),7.63 methoxybenzamido)methyl)phenyl)- (dd, J = 9.2, 3.2 Hz, 1H), 7.58(d, J = 8.5 Hz, 2H), 8-oxo-7,8-dihydro-9H-purin-9- 7.47-7.37 (m, 2H),7.26 (dd, J = 7.6, 3.3 Hz, 1H), yl)bicyclo[2.2.2]octane-1-carboxylic7.23-7.15 (m, 1H), 4.71 (s, 2H), 3.99 (s, 3H), 2.62 acid (48) (dd, J =9.6, 6.4 Hz, 6H), 2.02 (dd, J = 9.5, 6.5 Hz, 6H).

Example 26.N-(4-(8-amino-3-(4-(hydroxymethyl)bicyclo[2.2.2]octan-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 35)

To a solution of 33 (100 mg, 0.18 mmol) in THE (20 mL) at 0° C. wasadded lithium aluminum hydride (7 mg, 0.18 mmol). After stirring for 1hour, the mixture was added with saturated brine (0.5 mL) and thesolvent was removed under reduced pressure. The residue was purified byTLC (dichloromethane/methanol=12/1) to give the title compound 35 (32.2mg, solid, 34%).

MS m/z (ESI): 530 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.79 (d, J=5.3 Hz, 1H), 7.62 (dd, J=9.2, 3.2Hz, 1H), 7.59-7.50 (m, 4H), 7.25 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.17(dd, J=9.1, 4.2 Hz, 1H), 6.92 (d, J=5.3 Hz, 1H), 4.69 (s, 2H), 3.96 (s,3H), 3.26 (s, 2H), 2.14 (dd, J=9.4, 6.3 Hz, 6H), 1.65-1.59 (m, 6H).

The compound as shown below was synthesized according to the proceduresin Example 26, except that a different ester was used instead of 33.

Compound replacing MS m/z Compound No. Structure 33 (ESI) 50

50a 521

The NMR data of Compound 50 is shown below:

Compound ¹H NMR N-(4-(6-amino-9-((1r,4r)-4- ¹H NMR (400 MHz, CD₃OD) δ8.15 (hydroxymethyl)cyclohexyl)- (s, 1H), 7.64 (dd, J = 9.2, 3.3 Hz,1H), 8-oxo-8,9-dihydro-7H- 7.60 (d, J = 8.4 Hz, 2H), 7.46 (d, J = 8.4purin-7-yl)benzyl)-5-fluoro- Hz, 2H), 7.28 (ddd, J = 9.1, 7.6, 3.3 Hz,2-methoxybenzamide (50) 1H), 7.19 (dd, J = 9.2, 4.2 Hz, 1H), 4.72 (s,2H), 4.45-4.32 (m, 1H), 3.99 (s, 3H), 3.44 (d, J = 6.4 Hz, 2H),2.53-2.43 (m, 2H), 2.00 (d, J = 13.9 Hz, 2H), 1.90 (d, J = 9.2 Hz, 2H),1.62 (s, 1H), 1.21-1.12 (m, 2H).

Example 27.4-(8-amino-1-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[1,5-a]pyrazin-3-yl)-N-methylbicyclo[2.2.2]octane-1-carboxamide(Compound 36)

To a mixture of 34 (82 mg, 0.15 mmol), methylamine hydrochloride (20 mg,0.3 mmol) and DIPEA (78 mg, 0.6 mmol) in DMF (1.5 mL) was added HATU(114 mg, 0.3 mmol). After stirring for 30 minutes, the mixture waspurified by prep-HPLC to give the title compound 36 (11.5 mg, solid,14%).

MS m/z (ESI): 557 [M+1]

1H NMR (400 MHz, CD₃OD) δ 7.83 (d, J=5.4 Hz, 1H), 7.70-7.50 (m, 5H),7.26 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.17 (dd, J=9.1, 4.2 Hz, 1H), 6.93(d, J=5.4 Hz, 1H), 4.70 (d, J=4.0 Hz, 2H), 3.97 (s, 3H), 2.72 (t, J=5.0Hz, 3H), 2.19 (dd, J=9.6, 6.0 Hz, 6H), 1.93 (dd, J=9.5, 6.1 Hz, 6H).

Example 28.N-(4-(8-amino-3-(4-(morpholine-4-carbonyl)bicyclo[2.2.2]octan-1-yl)imidazo[1,5-a]pyrazin-1-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 37)

To a mixture of 34 (82 mg, 0.15 mmol), morpholine (26 mg, 0.3 mmol) andDIPEA (78 mg, 0.6 mmol) in DMF (1.5 mL) was added HATU (114 mg, 0.3mmol). After stirring for 30 minutes, the mixture was purified byprep-HPLC to give the title compound 37 (17.2 mg, solid, 19%).

MS m/z (ESI): 613 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.84 (d, J=5.4 Hz, 1H), 7.62 (dd, J=9.2, 3.2Hz, 1H), 7.55 (dd, J=20.6, 8.3 Hz, 4H), 7.26 (ddd, J=9.1, 7.6, 3.3 Hz,1H), 7.17 (dd, J=9.1, 4.2 Hz, 1H), 6.93 (d, J=5.4 Hz, 1H), 4.69 (s, 2H),3.97 (s, 3H), 3.71 (d, J=5.0 Hz, 4H), 3.67 (d, J=4.9 Hz, 4H), 2.25-2.15(m, 6H), 2.08 (dd, J=9.7, 5.4 Hz, 6H).

The compound as shown below was synthesized according to the proceduresin Example 28, except that a different acid was used instead of 34.

Compound Compound replacing MS m/z No. Structure 34 (ESI) 54

48 630

The NMR data of compounds 54 are shown below:

Compound ¹H NMR N-(4-(6-amino-9-(4- ¹H NMR (400 MHz, CD₃OD) δ8.10 (s,(morpholine-4- 1H), 7.63 (dd, J = 9.2, 3.3 Hz, 1H), 7.59carbonyl)bicyclo[2.2.2]octan- (s, 1H), 7.57 (s, 1H), 7.43 (s, 1H), 7.411-yl)-8-oxo-8,9-dihydro- (s, 1H), 7.31-7.25 (m, 1H), 7.19 (dd, J =7H-purin-7-yl)benzyl)- 9.2, 4.3 Hz, 1H), 4.71 (s, 2H), 3.99 (s,5-fluoro-2- 3H), 3.70 (s, 4H), 3.67 (d, J = 4.8 Hz, methoxybenzamide(54) 4H), 2.64 (d, J = 8.4 Hz, 6H), 2.13-2.09 (m, 6H).

Example 29.N-(4-(4-amino-7-morpholinoimidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 45)

Step 1. methyl 1-amino-1H-imidazole-5-carboxylate (45b)

To a solution of methyl 1H-imidazole-5-carboxylate 45a (630 mg, 5.0mmol) in THE at −78° C. was added a solution of LiHMDS in THF (1 M, 5mL). The solution was warmed to −10° C., stirred for 3 hours and thenadded with diphenylphosphonylhydroxylamine (1.28 g, 5.5 mmol). Themixture was warmed to room temperature and stirred for additional 12hours. It was concentrated to dryness under reduced pressure and theresidue was purified by silica gel column chromatography(dichloromethane/methanol=10/1) to give the title compound 45b (450 mg,63%).

MS m/z (ESI): 142 [M+1]

Step 2. imidazo[5,1-f][1,2,4]triazin-4(3H)-one (45c)

A mixture of 45b (450 mg, 3.17 mmol) and formamidine acetate (1.65 g,15.85 mmol) in ethanol (10 mL) was heated to 85° C. and stirred for 2days. After cooling to room temperature, the mixture was concentrated todryness to which was added water (50 mL). The mixture was filtered, andthe filter cake was collected to give the title compound 45c (330 mg,76%).

MS m/z (ESI): 137 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 11.88 (s, 1H), 8.47 (d, J=0.7 Hz, 1H), 7.93(s, 1H), 7.79 (d, J=0.7 Hz, 1H).

Step 3. 5,7-dibromoimidazo[5,1-f][1,2,4]triazin-4(3H)-one (45d)

To a solution of 45c (450 mg, 3.3 mmol) in DMF (5 mL) at 0° C. was addedliquid bromine (1.6 g). After stirring for 3 hours, the mixture wasfiltered to give the title compound 45d (550 mg, 77%).

MS m/z (ESI): 293 [M+1]

Step 4. 5-bromo-7-morpholinoimidazo[5,1-f][1,2,4]triazin-4(3H)-one (45e)

A mixture of 45d (300 mg, 1.02 mmol), morpholine (89 mg, 1.02 mmol),DMSO (1 mL) and DIPEA (263 mg, 2.04 mmol) was heated to 95° C. andstirred for 12 hours. After cooling to room temperature, the mixture wasconcentrated to dryness. The residue was dispersed in water (5 mL) andextracted with ethyl acetate (2×10 mL). The combined organic phase waswashed with saturated brine, dried over anhydrous sodium sulfate,filtered, and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography(dichloromethane/methanol=10/1) to give the title compound 45e (220 mg,72%).

MS m/z (ESI): 300 [M+1]

Step 5. 5-bromo-7-morpholinoimidazo[5,1-f][1,2,4]triazin-4-amine (45f)

To a solution of 1H-[1,2,4]triazole (455 mg, 6.6 mmol) in pyridine (5mL) was added phosphine oxychloride (335 mg, 2.19 mmol). The mixture wasstirred for 10 minutes, and then cooled to 0° C. to which was added asolution of 45e (220 mg, 0.73 mmol) in pyridine (3 mL). After stirringfor 2 hours at 0° C., a solution of ammonia in isopropanol (2 M, 3.65mL) was added. The resulting mixture was stirred at room temperature for12 hours and concentrated to dryness under reduced pressure. The residuewas purified by silica gel column chromatography(dichloromethane/methanol=10/1) to give the title compound 45f (86.5 mg,59.3%).

MS m/z (ESI): 299 [M+1]

Step 6.N-(4-(4-amino-7-morpholinoimidazo[5,1-f][1,2,4]triazin-5-yl)benzyl)-5-fluoro-2-methoxybenzamide(45)

To a mixture of 45f (86.5 mg, 0.3 mmol), 1b (115.5 mg, 0.3 mmol),potassium carbonate (82.8 mg, 0.6 mmol), 1,4-dioxane (5 mL) and water (1mL) was added PdCl₂(dppf) (24.5 mg, 0.03 mmol). The mixture was heatedto 90° C. and stirred for 2 hours. After cooling to room temperature,the reaction mixture was concentrated to dryness. The residue wasdispersed in water (10 mL) and extracted with ethyl acetate (2×20 mL).The combined organic phase was washed with saturated brine (10 mL),dried over anhydrous sodium sulfate, filtered, and concentrated todryness under reduced pressure. The residue was purified by prep-HPLC togive the title compound 45 (10 mg, solid, 7%).

MS m/z (ESI): 478 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.70 (s, 1H), 7.66-7.58 (m, 3H), 7.52 (d,J=8.3 Hz, 2H), 7.26 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.17 (dd, J=9.1, 4.2Hz, 1H), 4.68 (s, 2H), 3.97 (s, 3H), 3.89-3.81 (m, 4H), 3.63-3.53 (m,4H).

Example 30.(1s,4s)-4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)-1,4-dimethylcyclohexane-1-carboxylicacid (Compound 46)

Step 1. dimethyl (1s,4s)-1,4-dimethylcyclohexane-1,4-dicarboxylate (46a)and dimethyl (1r,4r)-1,4-dimethylcyclohexane-1,4-dicarboxylate (46b)

To a solution of LDA (2 M, 62.5 mL, 125 mmol) and hexamethylphosphorictriamide (69.5 mL, 400 mmol) in THF (125 mL) at −50° C. was added asolution of 32a (10 g, 50 mmol) in 1,4-dioxane solution (10 mL)dropwise. After stirring for 1 hour, the solution was warmed to 0° C.and stirred for another hour. After cooling to −50° C. again,iodomethane (31 g, 218.4 mmol) was added to the solution and stirringwas continued at room temperature for 16 hours. The solution was addedwith 1 N hydrochloric acid (30 mL) and extracted with ethyl acetate (50mL). The combined organic phase was dried over anhydrous sodium sulfate,filtered and concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=78/22) to give the title compound 46a (2.9 g, 250%) and 46b (1.4g, 12%).

Step 2. (1s,4s)-4-(methoxycarbonyl)-1,4-dimethylcyclohexane-1-carboxylicacid (46c)

A mixture of 46a (1 g, 4.38 mmol), potassium hydroxide (334 mg, 6.0mmol), methanol (20 mL) and water (20 mL) was heated to 80° C. andstirred for 16 hours. After cooling to room temperature, the solutionwas concentrated to dryness. The residue was added with 1 N hydrochloricacid (20 mL) and extracted with ethyl acetate (30 mL). The combinedorganic phase was dried over anhydrous sodium sulfate, filtered andconcentrated in vacuo to give the title compound 46c (795 mg, 85%).

Step 3 to 8. methyl(1s,4s)-4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)-1,4-dimethylcyclohexane-1-carboxylate(46i)

46i was synthesized according to the procedures for the first to sixthstep in Example 18, except that 46c was used instead of(R)-1-((benzyloxy)carbonyl)pyrrolidine-3-carboxylic acid 22a in thethird step.

MS m/z (ESI): 561 [M+1]

Step 9.(1s,4s)-4-(4-amino-5-(4-((5-fluoro-2-methoxybenzamido)methyl)phenyl)imidazo[5,1-f][1,2,4]triazin-7-yl)-1,4-dimethylcyclohexane-1-carboxylicacid (46)

46 was synthesized according to the procedures for the first to thesixth step in Example 26, except that 46i was used instead of 33 in thethird step.

MS m/z (ESI): 547 [M+1]

¹H NMR (400 MHz, DMSO-d₆) δ 12.00 (s, 1H), 8.84 (t, J=6.0 Hz 1H), 7.87(s, 1H), 7.59 (d, J=8.2 Hz, 2H), 7.53 (m, 1H), 7.47 (d, J=8.2 Hz, 2H),7.34 (s, 1H), 7.19 (m, 1H), 4.57 (d, J=6.1 Hz, 2H), 3.90 (s, 3H), 3.29(s, 2H), 2.33 (s, 2H), 1.88 (m, 4H), 1.44 (m, 5H), 1.18 (s, 3H).

Compound 47 as shown below was synthesized according to the proceduresfor the second to the ninth step in Example 30, except that 46b was usedinstead of 46a in the second step.

Compound Compound replacing MS m/z No. Structure 46a (ESI) 47

547

The NMR data of Compound 47 is shown below:

Compound ¹H NMR (1r,4r)-4-(4-amino-5-(4- ¹H NMR (400 MHz, DMSO-d₆) δ((5-fluoro-2- 12.10 (s, 1H), 8.84 (t, J = 6.0 Hz, 1H), 7.89methoxybenzamido)meth- (s, 1H), 7.61 (d, J = 8.2 Hz, 2H), 7.53 (m,yl)phenyl)imidazo[5,1- 1H), 7.48 (d, J = 8.2 Hz, 2H), 7.37-7.31 (m,f][1,2,4]triazin-7-yl)-1,4- 1H), 7.19 (m, 1H), 4.57 (d, J = 6.0 Hz, 2H),dimethylcyclohexane-1- 3.90 (s, 3H), 3.31 (s, 2H), 2.75 (d, J = 13.1carboxylic acid (47) Hz, 2H), 1.93 (d, J = 13.6 Hz, 2H), 1.45 (t, J =11.7 Hz, 2H), 1.33 (m, 5H), 1.00 (s, 3H).

Example 31.N-(4-(6-amino-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-7-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 51)

Step 1. N,N-dibenzyl-6-chloro-5-nitropyrimidin-4-amine (51a)

To a mixture of 4,6-dichloro-5-nitropyrimidine 51a (1.93 g, 10 mmol) andtriethylamine (2.02 g, 20 mmol) in dichloromethane (10 mL) was added asolution of dibenzylamine (1.97 g, mmol) in dichloromethane (5 mL) over10 minutes. The resulting mixture was stirred at 0° C. for 1 hour andconcentrated to dryness under reduced pressure. The residue wassuspended in water (70 mL) and extracted with ethyl acetate (2×100 mL).The combined organic phase was washed with saturated brine (50 mL),dried over anhydrous sodium sulfate, filtered and concentrated todryness under reduced pressure. The residue was purified by silica gelcolumn chromatography (petroleum ether/ethyl acetate=100/0 to 9/1) togive the title compound 51b (3.21 g, 91%).

MS m/z (ESI): 355[M+1]

Step 2.N⁴,N⁴-dibenzyl-5-nitro-NY-(tetrahydro-2H-pyran-4-yl)pyrimidine-4,6-diamine(51c)

To a mixture of 51b (0.8 g, 2.25 mmol) and triethylamine (455 mg, 4.5mmol) in dichloromethane (5 mL) was added tetrahydro-2H-pyran-4-amine(227 mg, 2.25 mmol) in dichloromethane (3 mL). The resulting mixture wasstirred for 1 hour and then concentrated to dryness under reducedpressure. The residue was suspended in water (70 mL) and extracted withethyl acetate (2×50 mL). The combined organic phase was washed withsaturated brine (50 mL), dried over anhydrous sodium sulfate, filteredand concentrated to dryness under reduced pressure. The residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=100/0 to 4/1) to give the title compound 51c (0.68 g, 72%).

MS m/z (ESI): 420[M+1]

Step 3.N⁴,N⁴-dibenzyl-N-(tetrahydro-2H-pyran-4-yl)pyrimidine-4,5,6-triamine(51d)

To a mixture of 51c (0.68 g, 1.62 mmol), ammonium chloride (433 mg, 8.1mmol), ethyl acetate (10 mL) and water (10 mL) was added zinc powder(526 mg, 8.1 mmol). The resulting mixture was stirred for 6 hours andextracted with ethyl acetate (2×50 mL). The combined organic phase waswashed with saturated brine (50 mL), dried over anhydrous sodiumsulfate, filtrated, and concentrated to dryness under reduced pressure.The residue was purified by silica gel column chromatography (petroleumether/ethyl acetate=100/0 to 1/1) to give the title compound 51d (0.52g, 82%).

MS m/z (ESI): 390[M+1]

Step 4.6-(dibenzylamino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one(51e)

To a mixture of triphosgene (520 mg, 1.33 mmol) and triethylamine (404mg, 4 mmol) in THE (10 mL) was added a solution of 51d (520 mg, 1.33mmol) in THE (3 mL). The resulting mixture was stirred for 1 hour andthen concentrated to dryness under reduced pressure. The residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=100/0 to 1/1) to give the title compound 51e (0.35 g, 82%).

MS m/z (ESI): 416[M+1]

Step 5. 6-amino-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one(51f)

To a solution of 51e (350 mg, 0.84 mmol) in acetic acid (10 mL) wasadded palladium on carbon (10%, 100 mg). The resulting mixture wasstirred for 12 hours under a hydrogen atmosphere and then filtered. Thefiltrate was concentrated to dryness under reduced pressure to give thetitle compound 51f (165 mg, 83%).

MS m/z (ESI): 236[M+1]

Step 6.N-(4-(6-amino-8-oxo-9-(tetrahydro-2H-pyran-4-yl)-8,9-dihydro-7H-purin-7-yl)benzyl)-5-fluoro-2-methoxybenzamide(51)

To a mixture of 51f (248 mg, 1.05 mmol) and 1b (636 mg, 2.1 mmol) inacetonitrile (5 mL) were added Cu(OAc)₂ (381 mg, 2.1 mmol) andtriethylamine (530 mg, 5.25 mmol)). The resulting mixture was stirredunder an air atmosphere for 12 hours and concentrated to dryness underreduced pressure. The residue was purified by prep-HPLC to give thetitle compound 51 (85.3 mg, solid, 17%).

MS m/z (ESI): 493[M+1]

¹H NMR (400 MHz, CD₃OD) δ 8.16 (s, 1H), 7.67-7.57 (m, 3H), 7.51-7.42 (m,2H), 7.28 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.20 (d, J=4.2 Hz, 1H), 4.72(s, 2H), 4.64 (tt, J=12.6, 4.4 Hz, 1H), 4.15-4.05 (m, 2H), 3.99 (s, 3H),3.57 (d, J=1.3 Hz, 2H), 2.77 (qd, J=12.6, 4.7 Hz, 2H), 1.79 (s, 2H).

The compounds or intermediates as shown below were synthesized accordingto the procedures in Example 31, except that a different amine was usedinstead of tetrahydro-2H-pyran-4-amine in the second step.

Compound/ Compound replacing MS Intermediate tetrahydro-2H-pyran- m/zNo. Structure 4-amine (ESI) 48a

575 50a

549 53

505

The NMR data of Compound 47 is shown below:

Compound ¹H NMR N-(4-(6-amino-8-oxo-9-(1,1,1- ¹H NMR (400 MHz, CD₃OD) δ8.17 trifluoropropan-2-yl)-8,9- (s, 1H), 7.69-7.57 (m, 3H), 7.52-7.43dihydro-7H-purin-7-yl)benzyl)-5- (m, 2H), 7.32-7.24 (m, 1H), 7.19 (dd,fluoro-2-methoxybenzamide (53) J = 9.1, 4.2 Hz, 1H), 5.23 (s, 1H), 4.72(s, 2H), 3.99 (s, 3H), 1.93 (d, J = 7.4 Hz, 3H).

Compound 49 can be synthesized according to the procedures in Example32.

Example 32.N-(4-(4-amino-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-1,2-dihydro-3H-imidazo[4,5-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(Compound 52)

Step 1. 2-chloro-3-nitro-N-(tetrahydro-2H-pyran-4-yl)pyridin-4-amine(52b)

To a mixture of 2,4-dichloro-3-nitropyridine 52a (1.93 g, 10 mmol) andtriethylamine (1.31 g, 13 mmol) in DMF (12 mL) was addedtetrahydro-2H-pyran-4-amine (1.01 g, 10 mmol). The resulting mixture wasstirred for 1 hour and then concentrated to dryness under reducedpressure.

The residue was suspended in water (70 mL) and extracted with ethylacetate (2×100 mL). The combined organic phase was washed with saturatedbrine (50 mL)c dried over anhydrous sodium sulfate, filtered andconcentrated to dryness under reduced pressure. The residue was purifiedby silica gel column chromatography (petroleum ether/ethyl acetate=100/0to 9/1) to give the title compound 52b (2.33 g, 91%).

MS m/z (ESI): 258 [M+1]

Step 2.N²,N²-dibenzyl-3-nitro-N⁴-(tetrahydro-2H-pyran-4-yl)pyridine-2,4-diamine(52c)

To a mixture of 52b (2.33 g, 9.07 mmol) and triethylamine (1.83 g, 18.1mmol) in acetonitrile (10 mL) was added dibenzylamine (1.78 g, 9.07mmol). The resulting mixture was heated to 80° C. and stirred for 20minutes. After cooling to room temperature, the reaction mixture wasconcentrated to dryness under reduced pressure and the residue waspurified by silica gel column chromatography (petroleum ether/ethylacetate=100/0 to 4/1) to give the title compound 52c (3.22 g, 85%).

MS m/z (ESI): 419 [M+1]

Step 3 to 6.N-(4-(4-amino-2-oxo-1-(tetrahydro-2H-pyran-4-yl)-1,2-dihydro-3H-imidazo[4,5-c]pyridin-3-yl)benzyl)-5-fluoro-2-methoxybenzamide(52)

52 was synthesized according to the procedures for the third to thesixth step in Example 31.

MS m/z (ESI): 492 [M+1]

¹H NMR (400 MHz, CD₃OD) δ 7.80 (s, 1H), 7.69-7.57 (m, 3H), 7.51-7.43 (m,2H), 7.28 (ddd, J=9.1, 7.6, 3.3 Hz, 1H), 7.20 (d, J=4.2 Hz, 1H), 6.95(s, 1H), 4.73 (s, 2H), 4.54 (tt, J=12.4, 4.2 Hz, 1H), 4.20-4.07 (m, 2H),3.99 (s, 3H), 3.60 (d, J=1.6 Hz, 2H), 2.56 (qd, J=12.6, 4.7 Hz, 2H),1.82 (dd, J=12.5, 2.6 Hz, 2H).

Biological Experiments Example 33. BTK Activity Inhibition Assay

In vitro kinase assay was used to evaluate the effects of compounds ofthe present invention on BTK activity (Table 1).

The experimental method is summarized below:

By using the homogeneous time-resolved fluorescence (HTRF) kinasedetection kit (Cisbio, catalog number 62TK0PEC), the enzymatic activityof BTK is determined by detecting the phosphorylation level of thesubstrate in the kinase reaction. The reaction buffer contains theenzyme reaction buffer (1×) from the kit, 5 mM MgCl₂, 1 mM DTT, 10 nMSEB and 0.01% Tween-20; the kinase reaction solution containshuman-derived recombinant BTK protein (Carna Biosciences, Catalog No.08-180) diluted to 0.2 ng/μL with the reaction buffer; the substratereaction solution contains biotin-labeled tyrosine kinase substratediluted to 0.5 μM with the reaction buffer and 40 μM ATP; the detectionbuffer contains Eu³⁺-labeled cage antibody diluted to 0.05 ng/μL andstreptavidin-labeled XL665 antibody diluted to 31.25 nM with thereaction buffer; the test compound is dissolved and diluted to 100 μMwith DMSO, followed by a 4-fold serial dilution with DMSO to the lowestconcentration of 6.1 nM and finally 40-time dilution with the reactionbuffer for each concentration point. If the IC₅₀ value of the compoundis very low, the initial concentration of the compound is reduced.

Add 4 μL test compound solution and 2 μL kinase reaction solution to a384-well detection plate (Corning, catalog number 3674), mix well andincubate at room temperature for 15 minutes; add 4 μL substrate reactionsolution and incubate for 50 minutes; add 10 μL detection buffer, mixwell and stand for 60 minutes; detect signal at 620 nm and 665 nm usingan Envision plate reader (Perkin Elmer). The signal value (absorbance at665 nm/absorbance at 620 nm) is positively correlated with the degree ofphosphorylation of the substrate, thereby detecting the kinase activityof BTK. In this experiment, the group without BTK is the negativecontrol (100% inhibition) and the group with BTK but no compound is thepositive control (0% inhibition). The inhibition curve is plotted andthe corresponding IC₅₀ value of the test compound is calculated usingXLfit software (ID Business Solutions Ltd., UK).

Example 34. BTK C481S Activity Inhibition Assay

In vitro kinase assay was used to evaluate the effects of compounds ofthe present invention on BTK C481S activity (Table 1).

The experimental method is summarized below:

By using the homogeneous time-resolved fluorescence (HTRF) kinasedetection kit (Cisbio, catalog number 62TK0PEC), the enzymatic activityof BTK C481S is determined by detecting the phosphorylation level of thesubstrate in the kinase reaction. The reaction buffer contains theenzyme reaction buffer (1×) from the kit, 5 mM MgCl₂, 1 mM DTT, 10 nMSEB, and 0.01% Tween-20; the kinase reaction solution contains humanrecombinant BTK C481S protein (purified in-house) diluted to 1.5 ng/μLwith the reaction buffer; the substrate reaction solution containsbiotin-labeled tyrosine kinase substrate diluted to 0.5 μM with thereaction buffer and 35 μM ATP; the detection buffer containsEu³⁺-labeled cage antibody diluted to 0.05 ng/μL andstreptavidin-labeled XL665 antibody diluted to 31.25 nM with thereaction buffer; the test compound is dissolved and diluted to 100 μMwith DMSO, followed by a 4-fold serial dilution with DMSO to the lowestconcentration of 6.1 nM and finally 40-time dilution with the reactionbuffer for each concentration point. If the IC₅₀ value of the compoundis very low, the initial concentration of the compound is reduced.

Add 4 μL test compound solution and 2 μL kinase reaction solution to a384-well detection plate (Corning, catalog number 3674), mix well andincubate at room temperature for 15 minutes; add 4 μL substrate reactionsolution and incubate for 50 minutes; add 10 μL detection buffer, mixwell and stand for 60 minutes; detect signal at 620 nm and 665 nm usingan Envision plate reader (Perkin Elmer). The signal value (absorbance at665 nm/absorbance at 620 nm) is positively correlated with the degree ofphosphorylation of the substrate, thereby detecting the kinase activityof BTK C481S. In this experiment, the group without BTK is the negativecontrol (100% inhibition) and the group with BTK C481 S but no compoundis the positive control (0% inhibition). The inhibition curve is plottedand the corresponding IC₅₀ value of the test compound is calculatedusing XLfit software (TD Business Solutions Ltd., UK).

TABLE 1 Compound No. BTK IC₅₀ (nM) BTK C481S IC₅₀ (nM) 1. 17 3.5 2. 1.72.5 3. 3.2 4.6 4. 2.3 3.0 5. 4.6 2.6 6. 4.6 3.3 7. 3.4 1.7 8. 1.2 0.8 9.718 498 10. 14 3.6 11. 20 3.9 12. 3.0 13. 1.4 0.8 14. 3.2 2.9 15. 1.41.1 16. 1.8 1.1 17. 6.3 2.4 18. 0.7 0.8 19. 212 201 21. 1.7 1.8 22. 15536 23. 1.0 3.0 24. 3.9 2.5 25. 1.1 1.5 26. 3.8 11 27. 2.7 1.3 28. 1.02.2 29. 1.2 2.1 31. 0.6 3.0 32. 2.4 6.3 33. 0.8 1.8 34. 0.9 1.5 35. 0.61.5 36. 0.9 3.4 37. 1.3 1.9 38. 2.9 2.1 39. 3.3 0.6 40. 0.4 1.1 41. 2.65.3 42. 1.7 4.3 43. 2.7 5.4 44. 10 7.1 45. 9.4 2.6 46. 7.7 1.6 47. 1.31.4 48. 1.2 8.2 50. 2.4 20 51. 4.0 9.2 52. 7.2 30 53. 5.6 21 54. 5.8 6.1

The invention, and the manner and process of making and using it, arenow described in such full, clear, concise and exact terms as to enableany person skilled in the art to which it pertains, to make and use thesame. It is to be understood that the foregoing describes preferredembodiments of the present invention and that modifications may be madetherein without departing from the scope of the present invention as setforth in the claims. To particularly point out and distinctly claim thesubject matter regarded as invention, the following claims conclude thespecification.

What is claimed is:
 1. A compound represented by general formula (I), ora pharmaceutically acceptable salt, stable isotope derivative, or stereoisomer thereof:

wherein: A is

where → indicates that A is connected to the benzene ring, and

indicates that A is connected to E; Ring K is phenyl or pyridyl, wherephenyl and pyridyl are optionally substituted by one or moresubstituents selected from halogen, cyano, C₁₋₆ alkyl or —OR^(a); E isC₁₋₆ alkyl, C₃₋₁₀ cycloalkyl, 3-10 membered heterocyclyl, C₆₋₁₀ aryl or5-10 membered heteroaryl, where the alkyl, cycloalkyl, heterocyclyl,aryl and heteroaryl are optionally substituted by one or moresubstituents selected from D, halogen, cyano, —OR^(b), —NR^(b)R^(c),—COOR^(b), —C(O)R^(b), —C(O)NR^(b)R^(c), or R^(e); R^(e) is C₁₋₆ alkyl,C₃₋₁₀ cycloalkyl or 3-10 membered heterocyclyl, where the alkyl,cycloalkyl and heterocyclyl are optionally substituted by halogen,cyano, —OR^(b), —NR^(b)R^(c), —COOR^(b), —C(O)R^(b) or —C(O)NR^(b)R^(c);R¹ is H, halogen, —OR^(a) or C₁₋₆ alkyl; R^(a) is C₁₋₆ alkyl, where oneor more hydrogens of the alkyl are optionally substituted by D orfluorine; and R^(b) and R^(c) are each independently selected from H,C₁₋₆ alkyl, C₃₋₆ cycloalkyl, or 4-6 membered heterocyclyl, wherein thecompound is a CNS penetrant having a Kp, CSF value of at least 0.15. 2.The compound according to claim 1, or a pharmaceutically acceptablesalt, stable isotope derivative, or a stereo isomer thereof, wherein Eis a C₁₋₆ alkyl group, wherein one or more hydrogens of the alkyl groupare optionally substituted by D or fluorine.
 3. The compound accordingto claim 1, or a pharmaceutically acceptable salt, stable isotopederivative, or a stereo isomer thereof, wherein E is C₃₋₇ monocycliccycloalkyl or 4-8 membered monocyclic heterocyclyl containing N and/orO, the monocyclic cycloalkyl and the monocyclic heterocyclyl areoptionally substituted by one or more substituents selected from D,halogen, —OR^(b), —NR^(b)R^(c), —COOR^(b), —C(O)R^(b), —C(O)NR^(b)R^(c)or C₁₋₆ alkyl, where one or more hydrogens of the alkyl group arefurther optionally substituted by halogen, —OR^(b) or —NR^(b)R^(c);R^(b) and R^(c) are each independently selected from H, C₁₋₆ alkyl, C₃₋₆cycloalkyl or N and/or O containing 4-6-membered heterocyclyl.
 4. Thecompound according to claim 1, or a pharmaceutically acceptable salt,stable isotope derivative, or a stereo isomer thereof, wherein E iscyclopropanyl, cyclobutanyl, cyclopentyl, cyclohexane, oxetanyl,tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, pyrrolidinyl,piperidinyl or morpholinyl, the cycloalkyl and heterocyclic groups areoptionally selected from one or more D, fluorine, —OR^(b), —NR^(b)R^(c),—COOR^(b), —C(O)R^(b), —C(O)NR^(b)R^(c) or C₁₋₂ alkyl substituentsubstituted, wherein one or more hydrogens of the alkyl substituents arefurther optionally substituted by fluorine, —OH or —NH₂; R^(b) and R^(c)are each independently selected from H, C₁₋₂ alkyl, C₃₋₆ cycloalkyl, or4-6 membered heterocyclyl containing N and/or O (for example,morpholinyl).
 5. The compound according to claim 1, or apharmaceutically acceptable salt, stable isotope derivative, or a stereoisomer thereof, wherein E is a C₅₋₁₀ polycyclic cycloalkyl group or an0-containing 5-10 membered polycyclic heterocyclic group, the polycycliccycloalkyl and polycyclic heterocyclic groups are optionally substitutedby one or more substituents selected from D, halogen, —OR^(b),—NR^(b)R^(c), —COOR^(b), —C(O)R^(b), —C(O)NR^(b)R^(c) or C₁₋₆ alkylsubstituents, wherein one or more hydrogens of the alkyl substituentsare further optionally substituted by halogen, —OR^(b) or —NR^(b)R^(c);R^(b) and R^(c) are each independently selected from H, C₁₋₆ alkyl, C₃₋₆cycloalkyl or 4-6 membered heterocyclic group containing N and/or O. 6.The compound according to claim 1, or a pharmaceutically acceptablesalt, stable isotope derivative, or a stereo isomer thereof, wherein Eis

 G is H, fluorine, —OR^(b), —NR^(b)R^(c), —COOR^(b), —C(O)R^(b),—C(O)NR^(b)R^(c) or C₁₋₂ alkyl, wherein one or more hydrogens of thealkyl are optionally substituted by fluorine, —OH or —NH₂; R^(b) andR^(c) are each independently selected from H, C₁₋₂ alkyl, C₃₋₆cycloalkyl or 4-6 membered heterocyclic group containing N and/or O (forexample, morpholinyl).
 7. The compound according to claim 1, or apharmaceutically acceptable salt, stable isotope derivative, or a stereoisomer thereof, wherein ring K is a phenyl group, wherein the phenylgroup is optionally selected from halogen or —OR^(a) by one or twogroups.
 8. The compound according to claim 1, or a pharmaceuticallyacceptable salt, stable isotope derivative, or a stereo isomer thereof,wherein ring K is

R^(a) is C₁₋₂ alkyl, wherein one or more hydrogens of the alkyl groupare optionally substituted by D.
 9. The compound according to claim 1,or its pharmaceutically acceptable salt, stable isotope derivative andisomer thereof, wherein R¹ is H or fluorine.
 10. A compound having thefollowing structure, or its pharmaceutically acceptable salt, stableisotope derivative, and stereoisomer thereof:


11. The compound of claim 10, which is:


12. The compound of claim 10, which is:


13. A pharmaceutical composition comprising the compound according toclaim 10, or a pharmaceutically acceptable salt, stable isotopederivative, or a stereoisomer thereof, and a pharmaceutically acceptablecarrier.
 14. A method for preventing or treating related diseasesmediated by BTK or its C481 mutant, the method comprising administeringto a patient in need a therapeutically effective amount of the compoundaccording to claim 10 or a pharmaceutically acceptable salt, stableisotope derivative, or a stereoisomer thereof, wherein the diseasesmediated by BTK and its C481 mutant are cancer, lymphoma, leukemia, anautoimmune disease or an inflammatory disease.
 15. The method accordingto claim 14, wherein the disease is B-cell lymphoma, chronic lymphocyticleukemia, small lymphocytic lymphoma, mantle cell lymphoma,Waldenstrom's macroglobulinemia, marginal zone lymphoma, follicularlymphoma, central nervous system lymphoma, non-Hodgkin's lymphoma,diffuse large B-cell lymphoma, multiple myeloma, rheumatoid arthritis,systemic lupus erythematosus, multiple sclerosis, lupus nephritis,Sjogren's syndrome, IgG4-related diseases, idiopathic thrombocytopenicpurpura, immune thrombocytopenia, or pemphigus.